Patent application title: METHODS FOR IDENTIFICATION, AND COMPOUNDS USEFUL FOR THE TREATMENT OF DEGENERATIVE & INFLAMMATORY DISEASES
Reginald Brys (Korbeek-Dijle, BE)
Nick Vandeghinste (Duffel, BE)
Peter Herwig Maria Tomme (Gent, BE)
IPC8 Class: AC40B3004FI
Class name: Combinatorial chemistry technology: method, library, apparatus method of screening a library by measuring the ability to specifically bind a target molecule (e.g., antibody-antigen binding, receptor-ligand binding, etc.)
Publication date: 2011-05-19
Patent application number: 20110118140
The present invention relates to in vivo and in vitro methods, agents and
compound screening assays for inhibiting extra-cellular matrix
degradation, including joint degenerative inhibiting and/or
anti-inflammatory pharmaceutical compositions, and the use thereof in
treating and/or preventing a disease involving extra-cellular matrix
degradation in a subject.
1. A method for identifying a compound that inhibits extra-cellular
matrix (ECM) degradation, comprising contacting a compound with a
polypeptide comprising the amino acid sequence of SEQ ID NO: 39; and
measuring a compound-polypeptide property related to extra-cellular
matrix (ECM) degradation.
2. The method according to claim 1, wherein said polypeptide is in an in vitro cell-free preparation.
3. The method of claim 1, wherein said property is a binding affinity of said compound to said polypeptide.
4. The method according to claim 1, wherein said compound is selected from the group consisting of compounds of a commercially available screening library and compounds having binding affinity for a polypeptide comprising the amino acid sequence of SEQ ID NO: 39.
5. The method according to claim 2, wherein said compound is a peptide in a phage display library or an antibody fragment library.
6. The method of claim 3, further comprising the steps of selecting a compound that exhibits moderate or high binding affinity to said polypeptide; contacting a population of mammalian cells with said compound; and measuring a second compound-polypeptide property related to extra-cellular matrix (ECM) degradation.
7. The method of claim 6, wherein said second compound-polypeptide property is activity of an ECM-degrading protein.
8. The method of claim 6, wherein said second compound-polypeptide property is expression of an ECM-degrading protein.
9. The method of claim 7, wherein said ECM-degrading protein is a Matrix Metallo Proteinase (MMP).
10. The method of claim 9, wherein said MMP is selected from the group consisting of MMP1, MMP2, MMP3, MMP8, MMP9, MMP13 and MMP14.
11. The method of claim 10, wherein said MMP is MMP1.
12. The method of claim 7, wherein said ECM-degrading protein is Cathepsin K.
13. The method of claim 8, wherein said ECM-degrading protein is a Matrix Metallo Proteinase (MMP).
14. The method of claim 13, wherein said MMP is selected from the group consisting of MMP1, MMP2, MMP3, MMP8, MMP9, MMP13 and MMP14.
15. The method of claim 14, wherein said MMP is MMP1.
16. The method of claim 8, wherein said ECM-degrading protein is Cathepsin K.
CROSS-REFERENCE TO RELATED APPLICATIONS
 This application is a divisional of U.S. application Ser. No. 11/854,037, filed Sep. 12, 2007, which is a divisional of U.S. application Ser. No. 11/152,366, filed Jun. 14, 2005, now U.S. Pat. No. 7,306,923, which claims priority to U.S. Provisional Application No. 60/579,307, filed Jun. 14, 2004, the disclosures of which are incorporated herein by reference.
FIELD OF INVENTION
 The present invention relates to a methods for identifying compounds, and expression-inhibition agents, capable of inhibiting the expression of proteins involved in the pathway resulting in the degradation of extra-cellular matrix (ECM), which inhibition is useful in the prevention and treatment of joint degeneration and diseases involving such degradation and/or inflammation.
 Diseases involving the degradation of extra-cellular matrix include, but are not limited to, psoriatic arthritis, juvenile arthritis, early arthritis, reactive arthritis, osteoarthritis, ankylosing spondylitis. osteoporosis, muskulo skeletal diseases like tendinitis and periodontal disease, cancer metastasis, airway diseases (COPD, asthma), renal and liver fibrosis, cardio-vascular diseases like atherosclerosis and heart failure, and neurological diseases like neuroinflammation and multiple sclerosis. Diseases involving primarily joint degeneration include, but are not limited to, psoriatic arthritis, juvenile arthritis, early arthritis, reactive arthritis, osteoarthritis, ankylosing spondylitis.
 Rheumatoid arthritis (RA) is a chronic joint degenerative disease, characterized by inflammation and destruction of the joint structures. When the disease is unchecked, it leads to substantial disability and pain due to loss of joint functionality and even premature death. The aim of an RA therapy, therefore, is not to slow down the disease but to attain remission in order to stop the joint destruction. Besides the severity of the disease outcome, the high prevalence of RA (0.8% of the adults are affected worldwide) means a high socio-economic impact. (For reviews on RA, we refer to Smolen and Steiner (2003); Lee and Weinblatt (2001); Choy and Panayi (2001); O'Dell (2004) and Firestein (2003)).
 Although it is widely accepted that RA is an auto-immune disease, there is no consensus concerning the precise mechanisms driving the `initiation stage` of the disease. What is known is that the initial trigger(s) does mediate, in a predisposed host, a cascade of events that leads to the activation of various cell types (B-cells, T-cells, macrophages, fibroblasts, endothelial cells, dendritic cells and others). Concomitantly, an increased production of various cytokines is observed in the joints and tissues surrounding the joint (e.g. TNF-α, IL-6, IL-1, IL-15, IL-18 and others). When the disease progresses, the cellular activation and cytokine production cascade becomes self-perpetuating. At this early stage, the destruction of joint structures is already very clear at this early stage. Thirty percent of the patients have radiographic evidence of bony erosions at the time of diagnosis and this proportion increases to 60 percent after two years.
 Histologic analysis of the joints of RA patients clearly evidences the mechanisms involved in the RA-associated degradative processes. The synovium is a cell layer, composed of a sublining and a lining region that separates the joint capsule from the synovial cavity. The inflamed synovium is central to the pathophysiology of RA. Histological differences in the synovium between normal and RA patients are indicated in FIG. 1: A. The synovial joint is composed of two adjacent bony ends each covered with a layer of cartilage, separated by a joint space and surrounded by the synovial membrane and joint capsule. The synovial membrane is composed of the synovial lining (facing the cartilage and bone) which consists of a thin (1-3 cells) layer of synoviocytes and the sublining connective tissue layer that is highly vascularised. The synovial membrane covers almost all intra-articular structures except for cartilage. B. Like many other forms of arthritis, rheumatoid arthritis (RA) is initially characterized by an inflammatory response of the synovial membrane (`synovitis`) that is characterised by an important influx of various types of mononuclear cells as well as by the activation of the local or infiltrated mononuclear cells. The lining layer becomes hyperplastic (it can have a thickness of >20 cells) and the synovial membrane expands. However, in addition, the hallmark of RA is joint destruction: the joint spaces narrow or disappear as a sign of cartilage degradation and destructions of the adjacent bone, also termed `erosions`, have occurred. The destructive portion of the synovial membrane is termed `pannus`. Enzymes secreted by synoviocytes lead to cartilage degradation.
 This analysis shows that the main effector responsible for RA-associated joint degradation is the pannus, where the synovial fibroblast, by producing diverse proteolytic enzymes, is the prime driver of cartilage and bone erosion. In the advanced RA patient, the pannus mediates the degradation of the adjacent cartilage, leading to the narrowing of the joint space, and has the potential to invade adjacent bone and cartilage. As bone and cartilage tissues are composed mainly of collagen type I or II, respectively, the pannus destructive and invasive properties are mediated by the secretion of collagenolytic proteases, principally the matrix metallo proteinases (MMPs). The erosion of the bone under and adjacent to the cartilage is also part of the RA process, and results principally from the presence of osteoclasts at the interface of bone and pannus. Osteoclasts adhere to the bone tissue and form a closed compartment, within which the osteoclasts secrete proteases (Cathepsin K, MMP9) that degrade the bone tissue. The osteoclast population in the joint is abnormally increased by osteoblast formation from precursor cells induced by the secretion of the receptor activator of NFkB ligand (RANKL) by activated SFs and T-cells.
 Various collagen types have a key role in defining the stability of the extra-cellular matrix (ECM). Collagens type I and collagen type II, for example, are the main components of bone and cartilage, respectively. Collagen proteins typically organise into multimeric structures referred to as collagen fibrils. Native collagen fibrils are very resistant to proteolytic cleavage. Only a few types of ECM-degrading proteins have been reported to have the capacity to degrade native collagen: matrix-metallo proteases (MMPs) and Cathepsins. Among the Cathepsins, cathepsin K, which is active mainly in osteoclasts, is the best characterised. Among the MMPs, MMP1, MMP2, MMP8 MMP13 and MMP14 are known to have collagenolytic properties. The correlation between an increased expression of MMP1 by synovial fibroblasts (SFs) and the progression of the arthritic disease is well-established and is predictive for joint erosive processes (Cunnane et al., 2001). In the context of RA, therefore, MMP1 represents a highly relevant collagen degrading protein. In vitro, the treatment of cultured SFs with cytokines relevant in the RA pathology (e.g. TNF-α and IL1β) will increase the expression of MMP1 by these cells (Andreakos et al., 2003). Monitoring the levels of MMP1 expressed by SFs therefore is a relevant readout in the field of RA as it is indicative for the activation of SFs towards an erosive phenotype that, in vivo, is responsible for cartilage degradation Inhibition of the MMP1 expression by SFs represents a valuable therapeutic approach towards the treatment of RA.
 The activity of the ECM-degrading proteins can also be causative or correlate with the progression of various diseases different from RA, as e.g. other diseases that involve the degradation of the joints. These diseases include, but are not limited to, psoriatic arthritis, juvenile arthritis, early arthritis, reactive arthritis, osteo-arthritis, and ankylosing spondylitis. Other diseases that may be treatable with compounds identified according to the present invention and using the targets involved in the expression of MMPs as described herein are osteoporosis, muskulo skeletal diseases like tendinitis and periodontal disease (Gapski et al., 2004), cancer metastasis (Coussens et al., 2002), airway diseases (COPD, asthma) (Suzuki et al., 2004), lung, renal fibrosis (Schanstra et al., 2002), liver fibrosis associated with chronic hepatitis C (Reiff et al., 2005), cardio-vascular diseases like atherosclerosis and heart failure (Creemers et al., 2001), and neurological diseases like neuroinflammation and multiple sclerosis (Rosenberg, 2002). Patients suffering from such diseases may benefit from stabilizing the ECM (by protecting it from degradation).
 NSAIDS (Non-steroidal anti-inflammatory drugs) are used to reduce the pain associated with RA and improve life quality of the patients. These drugs will not, however, put a brake on the RA-associated joint destruction.
 Corticosteroids are found to decrease the progression of RA as detected radiographically and are used at low doses to treat part of the RA patients (30 to 60%). Serious side effects, however, are associated with long corticosteroid use (Skin thinning, osteoporosis, cataracts, hypertension, hyperlipidemia).
 Synthetic DMARDs (Disease-Modifying Anti-Rheumatic Drugs) (e.g. methotrexate, leflunomide, sulfasalazine) mainly tackle the immuno-inflammatory component of RA. As a main disadvantage, these drugs only have a limited efficacy (joint destruction is only slowed down but not blocked by DMARDs such that disease progression in the long term continues). The lack of efficacy is indicated by the fact that, on average, only 30% of the patients achieve a ACR50 score after 24 months treatment with methotrexate. This means that, according to the American College of Rheumatology, only 30% of the patients do achieve a 50% improvement of their symptoms (O'Dell et al., 1996). In addition, the precise mechanism of action of DMARDs is often unclear.
 Biological DMARDs (Infliximab, Etanercept, Adalimumab, Rituximab, CTLA4-Ig) are therapeutic proteins that do inactivate cytokines (e.g. TNF-α) or cells (e.g. T-cells or B-cells) that have an important role in the RA pathophysiology (Kremer et al., 2003; Edwards et al., 2004). Although the TNF-α-blockers (Infliximab, Etanercept, Adalimumab) and methotrexate combination therapy is the most effective RA treatment currently available, it is striking that even this therapy only achieves a 50% improvement (ACR50) in disease symptoms in 50-60% of patients after 12 months therapy (St Clair et al., 2004). Some adverse events warnings for anti-TNF-α drugs exist, shedding a light on the side effects associated to this type of drugs. Increased risk for infections (tuberculosis) hematologic events and demyelinating disorders have been described for the TNF-α blockers. (see also Gomez-Reino et al., 2003). Besides the serious side effects, the TNF-α blockers do also share the general disadvantages of the biologicals class of therapeutics, which are the unpleasant way of administration (frequent injections accompanied by infusion site reactions) and the high production cost. Newer agents in late development phase target T-cell co-stimulatory molecules and B-cells. The efficacy of these agents is expected to be similar to that of the TNF-α blockers. The fact that a variety of targeted therapies have similar but limited efficacies, suggests that there is a multiplicity of pathogenic factors for RA. This is also indicative for the deficiencies in our understanding of pathogenic events relevant to RA.
 The current therapies for RA are not satisfactory due to a limited efficacy (no adequate therapy exists for 30% of the patients). This calls for additional strategies to achieve remission. Remission is required since residual disease bears the risk of progressive joint damage and thus progressive disability. Inhibiting the immuno-inflammatory component of the RA disease, which represents the main target of drugs currently used for RA treatment, does not result in a blockade of joint degradation, the major hallmark of the disease.
 The histological analysis of RA patient joints clearly identifies the pannus, as an aggressive, invasive tissue that represents the main culprit in joint degradation. Within the pannus, the synovial fibroblasts represent a link between the initiation of the abnormally triggered immune system that lies at the basis of RA pathogenesis, and the ultimate joint erosion. As no current RA therapy efficiently abolishes the erosive activity of the pannus in the long term, the discovery of novel drugs and/or drug targets that inhibit the generation, and/or the activity, of the pannus would represent an important milestone for the development of novel RA treatments.
 The present invention is based on the discovery of that certain proteins function in the pathway that results in the expression of extra-cellular matrix (ECM) degradation proteases, such as MMP1, and that inhibitors of the activity of these proteins, are useful for the treatment of diseases involving the abnormally high expression of such proteases.
SUMMARY OF THE INVENTION
 The present invention relates to a method for identifying compounds that inhibit extra-cellular matrix (ECM) degradation, comprising contacting a compound with a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 27-51 (hereinafter "TARGETS") and fragments thereof under conditions that allow said polypeptide to bind to the compound, and measuring a compound-polypeptide property related to extra-cellular matrix (ECM) degradation.
 Aspects of the present method include the in vitro assay of compounds using polypeptide of a TARGET and fragments thereof including selected from the group consisting of SEQ ID NO. 232-295, and cellular assays wherein TARGET inhibition is followed by observing indicators of efficacy including, for example, TARGET expression levels and/or Matrix Metallo Proteinase-1 levels.
 The present invention also relates to expression inhibitory agents comprising a polynucleotide selected from the group of an antisense polynucleotide, a ribozyme, and a small interfering RNA (siRNA), wherein said polynucleotide comprises a nucleic acid sequence complementary to, or engineered from, a naturally occurring polynucleotide sequence encoding a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 27-51 and 232-295, pharmaceutical compositions comprising said agent, useful in the treatment, or prevention, of chronic joint degenerative diseases such as rheumatoid arthritis.
 Another aspect of the invention is a method of treatment, or prevention, of a condition involving extra-cellular matrix (ECM) degradation, in a subject suffering or susceptible thereto, by administering a pharmaceutical composition comprising an effective TARGET-expression inhibiting amount of a expression-inhibitory agent.
 A further aspect of the present invention is a method for diagnosis relating to disease conditions characterized by extra-cellular matrix (ECM) degradation comprising measurement of indicators of levels of TARGET expression in a subject.
 Another aspect of this invention relates to the use of the present compound in a therapeutic method, a pharmaceutical composition, and the manufacture of such composition, useful for the treatment of a disease involving inflammation, and in particular, a disease characteristic of abnormal matrix metallo proteases activity.
BRIEF DESCRIPTION OF THE DRAWINGS
 FIG. 1. Schematic view of a normal joint and its changes in rheumatoid arthritis (From Smolen and Steiner, 2003).
 FIG. 2. Characterization of the expression of MMP1 by synovial fibroblasts. In panel A, the MMP1 mRNA levels present in the SF lysate are determined by real-time PCR. These MMP1 levels are normalized to the 18S levels that are also determined by real-time PCR for the same samples. Panel B shows the MMP1 signal detected from the supernatant that is subjected to Western blotting for detection of MMP1 protein levels using an MMP1-specific polyclonal antibody. Panel C shows the results of subjecting the supernatant to a commercially available MMP1 "activity ELISA" (Amersham Biosciences). The signal represented is proportional to the MMP1 activity present in the samples tested.
 FIG. 3. Increased expression of MMP1 by SFs triggered with various model adenoviruses. The SF supernatant uninfected SFs and SFs infected with the indicated model recombinant adenoviruses is subjected to the MMP1 ELISA and the MMP1 level measured by using a luminescence generating substrate is shown.
 FIG. 4A. Layout of the 384 control plate produced for the MMP1 ELISA assay.
 FIG. 4B. A representative example of the performance of the control plate tested with the protocol described in Example 2.
 FIG. 5. Representative example of the performance of the MMP1 ELISA run on a subset of 384 Ad-cDNAs of the FlexSelect collection that are tested in duplicate in a primary screen (A) and a rescreen (B).
 FIG. 6. Downscaling of the collagen degradation assay.
 FIG. 7. Matching of the collagen degradation assay readout to the visual assessment of collagen degradation.
 FIG. 8. Comparison of the degradation of FITC-labeled collagen type II and FITC-labeled Collagen type I in the collagen degradation assay.
 FIG. 9. Performance of the collagen degradation assay.
 FIG. 10. Activation of SFs by various complex cytokine mixtures. Shown are the raw luminescence signals from MMP1 ELISA measurements of the supernatant of SFs collected 72 hours after being triggered with the indicated recombinant cytokines or with the supernatant of THP1 cells activated with the indicated cytokines These measurements are proportional to MMP1 levels.
 FIG. 11. Inhibition of the response of SFs to a complex cytokine mixture by two inhibitors.
 FIG. 12. Ad-siRNA medicated reduction in the expression of various target genes in SF's reduces the capacity of these cells to express MMPI as a response to cytokines A) Results of cells infected with 3, 7.5, 12 or 15 μL of Ad-siRNAs designed against GPR21, FZD4, TM7SF1, PGPEP1, SEPT1, CD72 and FXYD5; B) results of cells infected with 3, 6, 9, 12 and 15 μl of Ad-siRNAs designed against PRKCE, CAMK4 and MAPKAPK5; C) results of cells infected with 3, 6, 9, and 12 μL of Ad-siRNAs designed against RIPK2 and RIT1; and D) results of cells infected with 3, 6, 9, and 12 μL of Ad-sirNA's designed against PPST1, USP21 and STK24.
 FIG. 13. Effect of adenovirus-mediated overexpression of target genes in SFs on the MMP1 expression by these cells. A) Result of infection of SFs with recombinant adenoviruses driving the expression of SEPT1, TPST1, USP21, MKNK1 and RIPK2; B) result of infection of SFs with recombinant adenoviruses driving the expression of PGPEP1 and RIT1; C) result of infection of SFs with recombinant adenoviruses driving the expression of CAMK4, MST3 and PRKCE; and D) result of infection of SFs with recombinant adenoviruses driving the expression of CD72, TM7SF1 and GPR21.
 FIG. 14. Reduction, at the protein level, of the expression of MAPKAPK5, PRKCE and CAMK4 by infection of the cells with various Ad-siRNA viruses targeting these genes
 FIG. 15. Inhibition of the collagen degradation by SFs as a response to a complex cytokine mixture by infection of the cells with various "knock down" viruses.
 FIG. 16. Structure of short-hairpin RNA (shRNA) targeted against Homo sapiens receptor-interacting serine-threonine kinase 2 (RIPK2) mRNA.
 The following terms are intended to have the meanings presented therewith below and are useful in understanding the description and intended scope of the present invention.
 The term "agent" means any molecule, including polypeptides, polynucleotides and small molecules.
 The term "agonist" refers to a ligand that stimulates the receptor the ligand binds to in the broadest sense.
 The term "assay" means any process used to measure a specific property of a compound. A "screening assay" means a process used to characterize or select compounds based upon their activity from a collection of compounds.
 The term "binding affinity" is a property that describes how strongly two or more compounds associate with each other in a non-covalent relationship. Binding affinities can be characterized qualitatively, (such as "strong", "weak", "high", or "low") or quantitatively (such as measuring the KD).
 The term "carrier" means a non-toxic material used in the formulation of pharmaceutical compositions to provide a medium, bulk and/or useable form to a pharmaceutical composition. A carrier may comprise one or more of such materials such as an excipient, stabilizer, or an aqueous pH buffered solution. Examples of physiologically acceptable carriers include aqueous or solid buffer ingredients including phosphate, citrate, and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptide; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and/or nonionic surfactants such as TWEEN®, polyethylene glycol (PEG), and PLURONICS®.
 The term "complex" means the entity created when two or more compounds bind to each other.
 The term "compound" is used herein in the context of a "test compound" or a "drug candidate compound" described in connection with the assays of the present invention. As such, these compounds comprise organic or inorganic compounds, derived synthetically or from natural sources. The compounds include inorganic or organic compounds such as polynucleotides, lipids or hormone analogs that are characterized by relatively low molecular weights. Other biopolymeric organic test compounds include peptides comprising from about 2 to about 40 amino acids and larger polypeptides comprising from about 40 to about 500 amino acids, such as antibodies or antibody conjugates.
 The term "condition" or "disease" means the overt presentation of symptoms (i.e., illness) or the manifestation of abnormal clinical indicators (e.g., biochemical indicators). Alternatively, the term "disease" refers to a genetic or environmental risk of or propensity for developing such symptoms or abnormal clinical indicators.
 The term "contact" or "contacting" means bringing at least two moieties together, whether in an in vitro system or an in vivo system.
 The term "derivatives of a polypeptide" relates to those peptides, oligopeptides, polypeptides, proteins and enzymes that comprise a stretch of contiguous amino acid residues of the polypeptide and that retain the biological activity of the protein, e.g. polypeptides that have amino acid mutations compared to the amino acid sequence of a naturally-occurring form of the polypeptide. A derivative may further comprise additional naturally occurring, altered, glycosylated, acylated or non-naturally occurring amino acid residues compared to the amino acid sequence of a naturally occurring form of the polypeptide. It may also contain one or more non-amino acid substituents compared to the amino acid sequence of a naturally occurring form of the polypeptide, for example a reporter molecule or other ligand, covalently or non-covalently bound to the amino acid sequence.
 The term "derivatives of a polynucleotide" relates to DNA-molecules, RNA-molecules, and oligonucleotides that comprise a stretch or nucleic acid residues of the polynucleotide, e.g. polynucleotides that may have nucleic acid mutations as compared to the nucleic acid sequence of a naturally occurring form of the polynucleotide. A derivative may further comprise nucleic acids with modified backbones such as PNA, polysiloxane, and 2'-O-(2-methoxy)ethyl-phosphorothioate, non-naturally occurring nucleic acid residues, or one or more nuclei acid substituents, such as methyl-, thio-, sulphate, benzoyl-, phenyl-, amino-, propyl-, chloro-, and methanocarbanucleosides, or a reporter molecule to facilitate its detection.
 The terms "ECM-degrading protein" and "ECM-degrading activity" refer to a protein and activity, respectively, that is capable of degrading extra-cellular matrixes found in bone and cartilage.
 The term "effective amount" or "therapeutically effective amount" means that amount of a compound or agent that will elicit the biological or medical response of a subject that is being sought by a medical doctor or other clinician.
 The term "endogenous" shall mean a material that a mammal naturally produces. Endogenous in reference to the term "protease", "kinase", or G-Protein Coupled Receptor ("GPCR") shall mean that which is naturally produced by a mammal (for example, and not limitation, a human). In contrast, the term non-endogenous in this context shall mean that which is not naturally produced by a mammal (for example, and not limitation, a human). Both terms can be utilized to describe both "in vivo" and "in vitro" systems. For example, and not a limitation, in a screening approach, the endogenous or non-endogenous TARGET may be in reference to an in vitro screening system. As a further example and not limitation, where the genome of a mammal has been manipulated to include a non-endogenous TARGET, screening of a candidate compound by means of an in vivo system is viable.
 The term "expressible nucleic acid" means a nucleic acid coding for a proteinaceous molecule, an RNA molecule, or a DNA molecule.
 The term "expression" comprises both endogenous expression and overexpression by transduction.
 The term "expression inhibitory agent" means a polynucleotide designed to interfere selectively with the transcription, translation and/or expression of a specific polypeptide or protein normally expressed within a cell. More particularly, "expression inhibitory agent" comprises a DNA or RNA molecule that contains a nucleotide sequence identical to or complementary to at least about 17 sequential nucleotides within the polyribonucleotide sequence coding for a specific polypeptide or protein. Exemplary expression inhibitory molecules include ribozymes, double stranded siRNA molecules, self-complementary single-stranded siRNA molecules, genetic antisense constructs, and synthetic RNA antisense molecules with modified stabilized backbones.
 The term "expressible nucleic acid" means a nucleic acid coding for a proteinaceous molecule, an RNA molecule, or a DNA molecule.
 The term "fragment of a polynucleotide" relates to oligonucleotides that comprise a stretch of contiguous nucleic acid residues that exhibit substantially a similar, but not necessarily identical, activity as the complete sequence.
 The term "fragment of a polypeptide" relates to peptides, oligopeptides, polypeptides, proteins and enzymes that comprise a stretch of contiguous amino acid residues, and exhibit substantially a similar, but not necessarily identical, functional activity as the complete sequence.
 The term "hybridization" means any process by which a strand of nucleic acid binds with a complementary strand through base pairing. The term "hybridization complex" refers to a complex formed between two nucleic acid sequences by virtue of the formation of hydrogen bonds between complementary bases. A hybridization complex may be formed in solution (e.g., C0t or R0t analysis) or formed between one nucleic acid sequence present in solution and another nucleic acid sequence immobilized on a solid support (e.g., paper, membranes, filters, chips, pins or glass slides, or any other appropriate substrate to which cells or their nucleic acids have been fixed). The term "stringent conditions" refers to conditions that permit hybridization between polynucleotides and the claimed polynucleotides. Stringent conditions can be defined by salt concentration, the concentration of organic solvent, e.g., formamide, temperature, and other conditions well known in the art. In particular, reducing the concentration of salt, increasing the concentration of formamide, or raising the hybridization temperature can increase stringency.
 The term "inhibit" or "inhibiting", in relationship to the term "response" means that a response is decreased or prevented in the presence of a compound as opposed to in the absence of the compound.
 The term "inhibition" refers to the reduction, down regulation of a process or the elimination of a stimulus for a process that results in the absence or minimization of the expression of a protein or polypeptide.
 The term "induction" refers to the inducing, up-regulation, or stimulation of a process that results in the expression of a protein or polypeptide.
 The term "ligand" means an endogenous, naturally occurring molecule specific for an endogenous, naturally occurring receptor.
 The term "pharmaceutically acceptable salts" refers to the non-toxic, inorganic and organic acid addition salts, and base addition salts, of compounds of the present invention. These salts can be prepared in situ during the final isolation and purification of compounds useful in the present invention.
 The term "polypeptide" relates to proteins, proteinaceous molecules, fractions of proteins, peptides, oligopeptides, enzymes (such as kinases, proteases, GCPR's etc.).
 The term "polynucleotide" means a polynucleic acid, in single or double stranded form, and in the sense or antisense orientation, complementary polynucleic acids that hybridize to a particular polynucleic acid under stringent conditions, and polynucleotides that are homologous in at least about 60 percent of its base pairs, and more preferably 70 percent of its base pairs are in common, most preferably 90 percent, and in a special embodiment 100 percent of its base pairs. The polynucleotides include polyribonucleic acids, polydeoxyribonucleic acids, and synthetic analogues thereof. It also includes nucleic acids with modified backbones such as peptide nucleic acid (PNA), polysiloxane, and 2'-O-(2-methoxy)ethylphosphorothioate. The polynucleotides are described by sequences that vary in length, that range from about 10 to about 5000 bases, preferably about 100 to about 4000 bases, more preferably about 250 to about 2500 bases. One polynucleotide embodiment comprises from about 10 to about 30 bases in length. A special embodiment of polynucleotide is the polyribonucleotide of from about 10 to about 22 nucleotides, more commonly described as small interfering RNAs (siRNAs). Another special embodiment are nucleic acids with modified backbones such as peptide nucleic acid (PNA), polysiloxane, and 2'-O-(2-methoxy)ethylphosphorothioate, or including non-naturally occurring nucleic acid residues, or one or more nucleic acid substituents, such as methyl-, thio-, sulphate, benzoyl-, phenyl-, amino-, propyl-, chloro-, and methanocarbanucleosides, or a reporter molecule to facilitate its detection.
 The term "polypeptide" relates to proteins (such as TARGETS), proteinaceous molecules, fractions of proteins peptides and oligopeptides.
 The term "solvate" means a physical association of a compound useful in this invention with one or more solvent molecules. This physical association includes hydrogen bonding. In certain instances the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. "Solvate" encompasses both solution-phase and isolable solvates. Representative solvates include hydrates, ethanolates and methanolates.
 The term "subject" includes humans and other mammals. The term "TARGET" or "TARGETS" means the protein(s) identified in accordance with the present assay to be involved in the induction of MMP1 levels. The preferred TARGETS are identified as SEQ ID NOS. 27-51 in Table 1. The more preferred TARGETS are the kinases, proteases and G-Protein Coupled Receptors (GPCRs) identified in Table 1.
 "Therapeutically effective amount" means that amount of a drug or pharmaceutical agent that will elicit the biological or medical response of a subject that is being sought by a medical doctor or other clinician. In particular, with regard to treating an disease condition characterized by the degradation of extracellular matrix, the term "effective matrix metallo-protease inhibiting amount" is intended to mean that effective amount of an compound of the present invention that will bring about a biologically meaningful decrease in the production of MMP-1 in the subject's disease affected tissues such that extracellular matrix degradation is meaningfully reduced. A compound having matrix metallo-protease inhibiting properties or a "matrix metallo-protease inhibiting compound" means a compound that provided to a cell in effective amounts is able to cause a biologically meaningful decrease in the production of MMP-I in such cells.
 The term "treating" means an intervention performed with the intention of preventing the development or altering the pathology of, and thereby alleviating a disorder, disease or condition, including one or more symptoms of such disorder or condition. Accordingly, "treating" refers to both therapeutic treatment and prophylactic or preventative measures. Those in need of treating include those already with the disorder as well as those in which the disorder is to be prevented. The related term "treatment," as used herein, refers to the act of treating a disorder, symptom, disease or condition, as the term "treating" is defined above.
Applicants' Invention Based on TARGET Relationship to Extra-cellular Matrix Degradation
 As noted above, the present invention is based on the present inventors' discovery that the TARGET polypeptides are factors in the up-regulation and/or induction of extra-cellular matrix degradation. The activity of the ECM-degrading protein is believed to be causative and to correlate with the progression of various diseases associated with an increased degradation of the extra-cellular matrix, including diseases that involve the degradation of the joint.
 The present invention relates to a method for assaying for drug candidate compounds that inhibit extra-cellular matrix degradation, comprising contacting the compound with a polypeptide comprising an amino acid sequence of SEQ ID NO: 27-51 and 232-295 under conditions that allow said polypeptide to bind to the compound, and detecting the formation of a complex between the polypeptide and the compound. One preferred means of measuring the complex formation is to determine the binding affinity of said compound to said polypeptide.
 More particularly, the invention relates to a method for identifying an agent that inhibits extra-cellular matrix degradation, the method comprising further:  (a) contacting a population of mammalian cells with one or more compound that exhibits binding affinity for a TARGET polypeptide, and  (b) measuring a compound-polypeptide property related to extra-cellular matrix degradation.
 The compound-polypeptide property referred to above is related to the expression of the TARGET, and is a measurable phenomenon chosen by the person of ordinary skill in the art. The measurable property may be, e.g., the binding affinity for a peptide domain of the polypeptide TARGET such as for SEQ ID NO: 232-295, or the level of any one of a number of biochemical marker levels of extra-cellular matrix degradation. Extra-cellular matrix degradation can e.g. be measured by measuring the level of enzymes that are induced during the process, such as expression of a MMP and/or a Cathepsin polypeptide.
 In a preferred embodiment of the invention, the TARGET polypeptide comprises an amino acid knockdown (KD) sequence selected from the group consisting of SEQ ID No: 27-51 as listed in Table 1.
TABLE-US-00001 TABLE 1 Ref/SEQ SEQ ID Ref/SEQ SEQ Gene accession NO accession ID NO Protein SEQ ID KD Hit No. Name Description (DNA) DNA (Protein) Protein Class Target H31- RIPK2 Homo sapiens receptor-interacting NM_003821 1 NP_003812 27 Kinase 52-56 290 serine-threonine kinase 2 (RIPK2), 168-170 mRNA. H31- PRKCE Homo sapiens protein kinase C, NM_005400 2 NP_005391 28 Kinase 57-61 035 epsilon (PRKCE), mRNA. 167 H31- MST3 Homo sapiens kinase SK246 from SK246 3 29 Kinase 62-66 319 Manning et al., Science. 164 NM_003576 4 30 Kinase 62-66 164 H34- MAPKAPK5 Homo sapiens mitogen-activated NM_003668 5 NP_003659 31 Kinase 67-71 088 protein kinase-activated protein 156-161 kinase 5 (MAPKAPK5), transcript variant 1, mRNA. NM_139078 6 32 Kinase 72-76 156-161 H34- MKNK1 Homo sapiens MAP kinase- NM_003684 7 NP_003675 33 Kinase 77-81 087 interacting serine/threonine kinase 1 162-163 (MKNK1), mRNA. H31- CAMK4 Homo sapiens calcium/calmodulin- NM_001744 8 NP_001735 34 Kinase 82-86 031 dependent protein kinase IV 148 (CAMK4), mRNA. CAMK4 SK061 9 35 Kinase 87-91 171 H31- SEPT1 Homo sapiens septin 1 (SEPT1), NM_052838 10 NP_443070 36 Secreted 92-96 347 mRNA. H31- PGPEP1 Homo sapiens pyroglutamyl- NM_017712 11 37 Protease 92-96 450 peptidase I 165-166 H31- CD72 Homo sapiens CD72 antigen NM_001782 12 NP_001773 38 Secreted 97-101 351 (CD72), mRNA. H31- TPST1 Homo sapiens tyrosylprotein NM_003596 13 NP_003587 39 Enzyme 102-106 301 sulfotransferase 1 (TPST1), mRNA. 150, 173 H31- GPR21 Homo sapiens G protein-coupled NM_005294 14 NP_005285 40 GPCR 107-111 242 receptor 21 (GPR21), mRNA. 155 H31- USP21 Homo sapiens ubiquitin specific NM_012475 15 NP_036607 41 Protease 112-116 047 protease 21 (USP21), transcript 174-175 variant 1, mRNA. USP21 Homo sapiens ubiquitin specific NM_016572 16 NP_057656 42 Protease 114-118 protease 21 (USP21), transcript 174-175 variant 2, mRNA. H34- FZD4 Homo sapiens frizzled homolog 4 NM_012193 17 NP_036325; 43 GPCR 119-123 092 (Drosophila) (FZD4), mRNA. 152-154 GAL_GPCR0379 18 GAL_GPCR_0379 44 GPCR 119-123 H31- TM7SF1 Homo sapiens transmembrane 7 NM_003272 19 NP_003263 45 GPCR 124-128 180 superfamily member 1 (upregulated 172 in kidney) (TM7SF1), mRNA. H31- FXYD5 Homo sapiens FXYD domain NM_014164 20 NP_054883 46 Secreted 129-133 384 containing ion transport regulator 5 151 (FXYD5), mRNA. H31- RIT1 Homo sapiens Ras-like without NM_006912 21 NP_008843 47 Enzyme 134-138 360 CAAX 1 (RIT1), mRNA H31- CASP10 Homo sapiens caspase 10, apoptosis- NM_001230 22 NP_001221 48 Protease 139-143 049 related cysteine protease (CASP10), 146 transcript variant A, mRNA. CASP10 Homo sapiens caspase 10, apoptosis- NM_032974 23 NP_116756 49 Protease 140-141 related cysteine protease (CASP10), 143-146 transcript variant B, mRNA. 149 CASP10 Homo sapiens caspase 10, apoptosis- NM_032976 24 NP_116758 50 Enzyme 139-143 related cysteine protease (CASP10), 146 transcript variant C, mRNA. CASP10 Homo sapiens caspase 10, apoptosis- NM_032977 25 NP_116759 51 Protease 140-143 related cysteine protease (CASP10), 146, 149 transcript variant D, mRNA. loop 26
 Depending on the choice of the skilled artisan, the present assay method may be designed to function as a series of measurements, each of which is designed to determine whether the drug candidate compound is indeed acting on the polypeptide to thereby inhibit extra-cellular matrix degradation. For example, an assay designed to determine the binding affinity of a compound to the polypeptide, or fragment thereof, may be necessary, but not sufficient, to ascertain whether the test compound would be useful for inhibiting extra-cellular matrix degradation when administered to a subject.
 Such binding information would be useful in identifying a set of test compounds for use in an assay that would measure a different property, further down the biochemical pathway, such as for example MMP-1 expression. Such second assay may be designed to confirm that the test compound, having binding affinity for the polypeptide, actually inhibits extra-cellular matrix degradation. Suitable controls should always be in place to insure against false positive readings.
 The order of taking these measurements is not believed to be critical to the practice of the present invention, which may be practiced in any order. For example, one may first perform a screening assay of a set of compounds for which no information is known respecting the compounds' binding affinity for the polypeptide. Alternatively, one may screen a set of compounds identified as having binding affinity for a polypeptide domain, or a class of compounds identified as being an inhibitor of the polypeptide. However, for the present assay to be meaningful to the ultimate use of the drug candidate compounds, a measurement of extra-cellular matrix degradation activity is necessary. Validation studies including controls, and measurements of binding affinity to the polypeptides of the invention are nonetheless useful in identifying a compound useful in any therapeutic or diagnostic application.
 The present assay method may be practiced in vitro, using one or more of the TARGET proteins, or fragments thereof. The amino acid sequences of exemplary protein domain fragments of selected TARGETS are SEQ ID NO: 232-295, listed in Table 1A below.
TABLE-US-00002 TABLE 1A SEQ ID NO Protein Accession Name Protein Segment segment NM_005294 GPR21 Extracellular domain 232 NM_005294 GPR21 Transmembrane domain 233 NM_005294 GPR21 Intracellular domain 234 NM_005294 GPR21 Transmembrane domain 235 NM_005294 GPR21 Extracellular domain 236 NM_005294 GPR21 Transmembrane domain 237 NM_005294 GPR21 Intracellular domain 238 NM_005294 GPR21 Transmembrane domain 239 NM_005294 GPR21 Extracellular domain 240 NM_005294 GPR21 Transmembrane domain 241 NM_005294 GPR21 Intracellular domain 242 NM_005294 GPR21 Transmembrane domain 243 NM_005294 GPR21 Extracellular domain 244 NM_005294 GPR21 Transmembrane domain 245 NM_005294 GPR21 Intracellular domain 246 NM_012193 FZD4 Extracellular domain 247 NM_012193 FZD4 Transmembrane domain 248 NM_012193 FZD4 Intracellular domain 249 NM_012193 FZD4 Transmembrane domain 250 NM_012193 FZD4 Extracellular domain 251 NM_012193 FZD4 Transmembrane domain 252 NM_012193 FZD4 Intracellular domain 253 NM_012193 FZD4 Transmembrane domain 254 NM_012193 FZD4 Extracellular domain 255 NM_012193 FZD4 Transmembrane domain 256 NM_012193 FZD4 Intracellular domain 257 NM_012193 FZD4 Transmembrane domain 258 NM_012193 FZD4 Extracellular domain 259 NM_012193 FZD4 Transmembrane domain 260 NM_012193 FZD4 Intracellular domain 261 NM_003272 TM7SF1 Extracellular domain 262 NM_003272 TM7SF1 Transmembrane domain 263 NM_003272 TM7SF1 Intracellular domain 264 NM_003272 TM7SF1 Transmembrane domain 265 NM_003272 TM7SF1 Extracellular domain 266 NM_003272 TM7SF1 Transmembrane domain 267 NM_003272 TM7SF1 Intracellular domain 268 NM_003272 TM7SF1 Transmembrane domain 269 NM_003272 TM7SF1 Extracellular domain 270 NM_003272 TM7SF1 Transmembrane domain 271 NM_003272 TM7SF1 Intracellular domain 272 NM_003272 TM7SF1 Transmembrane domain 273 NM_003272 TM7SF1 Extracellular domain 274 NM_003272 TM7SF1 Transmembrane domain 275 NM_003272 TM7SF1 Intracellular domain 276 NM_001782 CD72 Intracellular domain 277 NM_001782 CD72 Transmembrane domain 278 NM_001782 CD72 Extracellular domain 279 NM_014164 FXYD5 Extracellular domain 280 NM_014164 FXYD5 Transmembrane domain 281 NM_014164 FXYD5 Intracellular domain 282 GAL_GPCR0379 FZD4 Intracellular domain 283 GAL_GPCR0379 FZD4 Transmembrane domain 284 GAL_GPCR0379 FZD4 Extracellular domain 285 GAL_GPCR0379 FZD4 Transmembrane domain 286 GAL_GPCR0379 FZD4 Intracellular domain 287 GAL_GPCR0379 FZD4 Transmembrane domain 288 GAL_GPCR0379 FZD4 Extracellular domain 289 GAL_GPCR0379 FZD4 Transmembrane domain 290 GAL_GPCR0379 FZD4 Intracellular domain 291 GAL_GPCR0379 FZD4 Transmembrane domain 292 GAL_GPCR0379 FZD4 Extracellular domain 293 GAL_GPCR0379 FZD4 Transmembrane domain 294 GAL_GPCR0379 FZD4 Intracellular domain 295
 The binding affinity of a compound with the polypeptide TARGET can be measured by methods known in the art, such as using surface plasmon resonance biosensors (Biacore), by saturation binding analysis with a labeled compound (e.g. Scatchard and Lindmo analysis), by differential UV spectrophotometer, fluorescence polarization assay, Fluorometric Imaging Plate Reader (FLIPR®) system, Fluorescence resonance energy transfer, and Bioluminescence resonance energy transfer. The binding affinity of compounds can also be expressed in dissociation constant (Kd) or as 1050 or EC50. The 1050 represents the concentration of a compound that is required for 50% inhibition of binding of another ligand to the polypeptide. The EC50 represents the concentration required for obtaining 50% of the maximum effect in any assay that measures TARGET function. The dissociation constant, Kd, is a measure of how well a ligand binds to the polypeptide, it is equivalent to the ligand concentration required to saturate exactly half of the binding-sites on the polypeptide. Compounds with a high affinity binding have low Kd, 1050 and EC50 values, i.e. in the range of 100 nM to 1 pM; a moderate to low affinity binding relates to a high Kd, 1050 and EC50 values, i.e. in the micromolar range.
 The present assay method may also be practiced in a cellular assay, A host cell expressing the TARGET can be a cell with endogenous expression or a cell over-expressing the TARGET e.g. by transduction. When the endogenous expression of the polypeptide is not sufficient to determine a baseline that can easily be measured, one may use using host cells that over-express TARGET. Over-expression has the advantage that the level of the TARGET substrate end products is higher than the activity level by endogenous expression. Accordingly, measuring such levels using presently available techniques is easier.
 One embodiment of the present method for identifying a compound that decreases extra-cellular matrix (ECM) degradation comprises culturing a population of mammalian cells expressing a TARGET polypeptide, or a functional fragment or derivative thereof; determining a first level of ECM degradation in said population of cells; exposing said population of cells to a compound, or a mixture of compounds; determining a second level of ECM degradation in said population of cells during or after exposure of said population of cells to said compound, or the mixture of said compounds; and identifying the compound(s) that decreases ECM degradation. As noted above, ECM degradation may be determined by measuring the expression and/or activity of the TARGET polypeptide and/or a known ECM-degrading protein. In a preferred embodiment, said ECM-degrading protein is able to degrade collagen, and more preferably, is able to degrade collagen type I and/or collagen type II. In another preferred embodiment of the present invention, said ECM-degrading protein is a Matrix Metallo Proteinase (MMP), and more preferably is selected from the group consisting of: MMP1, MMP2, MMP3, MMP8, MMP9, MMP13 and MMP14. In this context, the most preferred ECM-degrading protein is Matrix Metalloprotease 1 (MMP1). In yet another preferred embodiment, said ECM-degrading protein is Cathepsin K.
 The expression of an ECM-degrading protein can be determined by methods known in the art such as Western blotting using specific antibodies, or an ELISA using antibodies specifically recognizing a particular ECM-degrading protein.
 The activity of an ECM-degrading protein can be determined by using fluorogenic small peptide substrates. The specificity of these substrates, however, is often limited. In general, the use of these substrates is limited to the testing of purified proteases in biochemical assays, to avoid interference of other proteases.
 The present inventors have developed a protocol allowing the detection, in a high throughput mode, of the activity of collagen degrading enzymes in complex media such as the supernatant of cultured cells. This protocol makes use of native collagen, being labelled with a fluorescent label, as a substrate.
 The present inventors identified target genes involved in ECM-degradation by using a `knock-in` library. This type of library is a screen in which cDNA molecules are transduced into cells by recombinant adenoviruses that induce the expression and activity of a specific gene and corresponding gene product in a cell. Each cDNA in a viral vector corresponds to a specific natural gene. By identifying a cDNA that stimulates ECM-degradation, a direct correlation between can be drawn between the specific gene expression and ECM degradation. The TARGET genes identified using the knock-in library (the protein expression products thereof herein referred to as "TARGET" polypeptides) are then used in the present inventive method for identifying compounds that can be used to prevent ECM-degradation. Indeed, shRNA compounds comprising the sequences listed in Table 3 (SEQ ID NO: 52-175) and the antisense sequences corresponding thereto inhibit the expression and/or activity of these TARGET genes and decrease the ECM-degrading activity of cells, confirming the role of these TARGET genes in ECM-degradation.
TABLE-US-00003 TABLE 3 List of target sequences selected within the coding sequences of the genes identified as modulators of the collagenolytic activity of SFs for use in RNAi-based down- regulation of the expression of these genes. DISPLAY_ID ACCESSION NAME SIRNA_NAME SEQ ID NO CAMK4 NM_001744 A150100-CAMK4_v1 NM_001744_idx445 83 CAMK4 NM_001744 A150100-CAMK4_v10 NM_001744_idx1045 148 CAMK4 NM_001744 A150100-CAMK4_v11 NM_001744_idx1186 85 CAMK4 NM_001744 A150100-CAMK4_v2 NM_001744_idx258 86 CAMK4 NM_001744 A150100-CAMK4_v3 NM_001744_idx668 84 CAMK4 NM_001744 A150100-CAMK4_v9 NM_001744_idx427 82 CASP10 NM_001230 A150100-CASP10_v1 NM_001230_idx934 146 CASP10 NM_001230 A150100-CASP10_v10 NM_001230_idx1532 142 CASP10 NM_001230 A150100-CASP10_v13 NM_001230_idx1111 143 CASP10 NM_001230 A150100-CASP10_v2 NM_001230_idx382 141 CASP10 NM_001230 A150100-CASP10_v8 NM_032974_idx317 140 CASP10 NM_032974 A150100-CASP10_v1 NM_001230_idx934 146 CASP10 NM_032974 A150100-CASP10_v11 NM_032974_idx1674 144 CASP10 NM_032974 A150100-CASP10_v12 NM_032974_idx1829 145 CASP10 NM_032974 A150100-CASP10_v13 NM_001230_idx1111 143 CASP10 NM_032974 A150100-CASP10_v2 NM_001230_idx382 141 CASP10 NM_032974 A150100-CASP10_v7 NM_032974_idx981 149 CASP10 NM_032974 A150100-CASP10_v8 NM_032974_idx317 140 CASP10 NM_032976 A150100-CASP10_v1 NM_001230_idx934 146 CASP10 NM_032976 A150100-CASP10_v10 NM_001230_idx1532 142 CASP10 NM_032976 A150100-CASP10_v13 NM_001230_idx1111 143 CASP10 NM_032976 A150100-CASP10_v2 NM_001230_idx382 141 CASP10 NM_032976 A150100-CASP10_v8 NM_032974_idx317 140 CASP10 NM_032977 A150100-CASP10_v1 NM_001230_idx934 146 CASP10 NM_032977 A150100-CASP10_v10 NM_001230_idx1532 142 CASP10 NM_032977 A150100-CASP10_v13 NM_001230_idx1111 143 CASP10 NM_032977 A150100-CASP10_v2 NM_001230_idx382 141 CASP10 NM_032977 A150100-CASP10_v7 NM_032974_idx981 149 CASP10 NM_032977 A150100-CASP10_v8 NM_032974_idx317 140 CD72 NM_001782 A150100-CD72_v2 NM_001782_idx376 100 CD72 NM_001782 A150100-CD72_v3 NM_001782_idx742 97 CD72 NM_001782 A150100-CD72_v4 NM_001782_idx975 150 CD72 NM_001782 A150100-CD72_v5 NM_001782_idx1049 98 CD72 NM_001782 A150100-CD72_v6 NM_001782_idx1054 101 CD72 NM_001782 A150100-CD72_v7 NM_001782_idx901 99 FXYD5 NM_014164 A150100-FXYD5_v2 NM_014164_idx224 132 FXYD5 NM_014164 A150100-FXYD5_v3 NM_014164_idx417 131 FXYD5 NM_014164 A150100-FXYD5_v4 NM_014164_idx436 129 FXYD5 NM_014164 A150100-FXYD5_v5 NM_014164_idx542 133 FXYD5 NM_014164 A150100-FXYD5_v6 NM_014164_idx603 130 FXYD5 NM_014164 A150100-FXYD5_v7 NM_014164_idx672 151 FZD4 NM_012193 A150100-C(27)-3BETA- NM_025193_idx1374 152 HSD_v3 FZD4 NM_012193 A150100-FZD4_v10 NM_012193_idx849 122 FZD4 NM_012193 A150100-FZD4_v5 NM_012193_idx481 120 FZD4 NM_012193 A150100-FZD4_v6 NM_012193_idx1570 153 FZD4 NM_012193 A150100-FZD4_v7 NM_012193_idx745 123 FZD4 NM_012193 A150100-FZD4_v8 NM_012193_idx1160 154 FZD4 NM_012193 A150100-FZD4_v9 NM_012193_idx534 121 GPR21 NM_005294 A150100-GPR21_v10 NM_005294_idx638 108 GPR21 NM_005294 A150100-GPR21_v11 NM_005294_idx936 109 GPR21 NM_005294 A150100-GPR21_v12 NM_005294_idx168 155 GPR21 NM_005294 A150100-GPR21_v13 NM_005294_idx868 107 GPR21 NM_005294 A150100-GPR21_v14 NM_005294_idx988 111 GPR21 NM_005294 A150100-GPR21_v9 NM_005294_idx161 110 MAPKAPK5 NM_003668 A150100- oKD102 70 MAPKAPK5_v1 MAPKAPK5 NM_003668 A150100- NM_003668_idx856 156 MAPKAPK5_v10 MAPKAPK5 NM_003668 A150100- NM_003668_idx1542 76 MAPKAPK5_v11 MAPKAPK5 NM_003668 A150100- NM_003668_idx456 157 MAPKAPK5_v12 MAPKAPK5 NM_003668 A150100- NM_003668_idx609 158 MAPKAPK5_v13 MAPKAPK5 NM_003668 A150100- oKD103 159 MAPKAPK5_v2 MAPKAPK5 NM_003668 A150100- oKD104 160 MAPKAPK5_v8 MAPKAPK5 NM_003668 A150100- NM_003668_idx686 161 MAPKAPK5_v9 MAPKAPK5 NM_139078 A150100- oKD102 70 MAPKAPK5_v1 MAPKAPK5 NM_139078 A150100- NM_003668_idx856 156 MAPKAPK5_v10 MAPKAPK5 NM_139078 A150100- NM_003668_idx1542 76 MAPKAPK5_v11 MAPKAPK5 NM_139078 A150100- NM_003668_idx456 157 MAPKAPK5_v12 MAPKAPK5 NM_139078 A150100- NM_003668_idx609 158 MAPKAPK5_v13 MAPKAPK5 NM_139078 A150100- oKD103 159 MAPKAPK5_v2 MAPKAPK5 NM_139078 A150100- oKD104 160 MAPKAPK5_v8 MAPKAPK5 NM_139078 A150100- NM_003668_idx686 161 MAPKAPK5_v9 MKNK1 NM_003684 A150100-MKNK1_v1 oKD110 162 MKNK1 NM_003684 A150100-MKNK1_v14 oKD109 81 MKNK1 NM_003684 A150100-MKNK1_v15 oKD108 77 MKNK1 NM_003684 A150100-MKNK1_v16 NM_003684_idx384 79 MKNK1 NM_003684 A150100-MKNK1_v17 NM_003684_idx549 80 MKNK1 NM_003684 A150100-MKNK1_v18 NM_003684_idx1216 163 MST3 SK246 A150100-MST3_v2 SK246_idx413 66 MST3 SK246 A150100-MST3_v3 SK246_idx508 65 MST3 SK246 A150100-MST3_v4 SK246_idx918 63 MST3 SK246 A150100-STK24_v1 NM_003576_idx300 62 MST3 SK246 A150100-STK24_v2 NM_003576_idx950 164 MST3 SK246 A150100-STK24_v3 NM_003576_idx1020 64 PGPEP1 NM_017712 A150100- NM_017712_idx176 94 FLJ20208_v10 PGPEP1 NM_017712 A150100- NM_017712_idx404 92 FLJ20208_v11 PGPEP1 NM_017712 A150100-FLJ20208_v5 NM_017712_idx289 96 PGPEP1 NM_017712 A150100-FLJ20208_v6 NM_017712_idx164 93 PGPEP1 NM_017712 A150100-FLJ20208_v7 NM_017712_idx496 165 PGPEP1 NM_017712 A150100-FLJ20208_v8 NM_017712_idx198 95 PGPEP1 NM_017712 A150100-FLJ20208_v9 NM_017712_idx298 166 PRKCE NM_005400 A150100-PRKCE_v10 NM_005400_idx760 59 PRKCE NM_005400 A150100-PRKCE_v11 NM_005400_idx1276 60 PRKCE NM_005400 A150100-PRKCE_v2 NM_005400_idx1240 57 PRKCE NM_005400 A150100-PRKCE_v7 NM_005400_idx1109 58 PRKCE NM_005400 A150100-PRKCE_v8 NM_005400_idx2050 61 PRKCE NM_005400 A150100-PRKCE_v9 NM_005400_idx148 167 RIPK2 NM_003821 A150100-RIPK2_v1 oKD111 52 RIPK2 NM_003821 A150100-RIPK2_v10 NM_003821_idx993 168 RIPK2 NM_003821 A150100-RIPK2_v11 NM_003821_idx1416 169 RIPK2 NM_003821 A150100-RIPK2_v2 oKD112 54 RIPK2 NM_003821 A150100-RIPK2_v3 oKD113 55 RIPK2 NM_003821 A150100-RIPK2_v9 NM_003821_idx612 170 RIT1 NM_006912 A150100-RIT_v2 NM_006912_idx247 137 RIT1 NM_006912 A150100-RIT_v3 NM_006912_idx536 134 RIT1 NM_006912 A150100-RIT_v4 NM_006912_idx622 136 RIT1 NM_006912 A150100-RIT_v5 NM_006912_idx824 138 RIT1 NM_006912 A150100-RIT_v6 NM_006912_idx263 135 SEPT1 NM_052838 A150100-SEPT1_v2 NM_052838_idx305 171 SEPT1 NM_052838 A150100-SEPT1_v3 NM_052838_idx329 89 SEPT1 NM_052838 A150100-SEPT1_v4 NM_052838_idx480 90 SEPT1 NM_052838 A150100-SEPT1_v5 NM_052838_idx677 88 SEPT1 NM_052838 A150100-SEPT1_v6 NM_052838_idx954 87 SEPT1 NM_052838 A150100-SEPT1_v7 NM_052838_idx1218 91 MST3 NM_003576 A150100-MST3_v2 SK246_idx413 66 MST3 NM_003576 A150100-MST3_v3 SK246_idx508 65 MST3 NM_003576 A150100-MST3_v4 SK246_idx918 63 MST3 NM_003576 A150100-STK24_v1 NM_003576_idx300 62 MST3 NM_003576 A150100-STK24_v2 NM_003576_idx950 164 MST3 NM_003576 A150100-STK24_v3 NM_003576_idx1020 64 TM7SF1 NM_003272 A150100-TM7SF1_v11 NM_003272_idx637 128 TM7SF1 NM_003272 A150100-TM7SF1_v12 NM_003272_idx673 125 TM7SF1 NM_003272 A150100-TM7SF1_v13 NM_003272_idx764 172 TM7SF1 NM_003272 A150100-TM7SF1_v14 NM_003272_idx775 127 TM7SF1 NM_003272 A150100-TM7SF1_v9 NM_003272_idx275 124 TPST1 NM_003596 A150100-TPST1_v1 NM_003596_idx722 106 TPST1 NM_003596 A150100-TPST1_v2 NM_003596_idx1262 104 TPST1 NM_003596 A150100-TPST1_v3 NM_003596_idx425 102 TPST1 NM_003596 A150100-TPST1_v5 NM_003596_idx1229 103 TPST1 NM_003596 A150100-TPST1_v6 NM_003596_idx1260 105 TPST1 NM_003596 A150100-TPST1_v7 NM_003596_idx1444 173 USP21 NM_012475 A150100-USP21_v1 NM_012475_idx1574 112 USP21 NM_012475 A150100-USP21_v13 NM_012475_idx741 117 USP21 NM_012475 A150100-USP21_v14 NM_012475_idx928 174 USP21 NM_012475 A150100-USP21_v15 NM_012475_idx682 114 USP21 NM_012475 A150100-USP21_v16 NM_012475_idx733 118 USP21 NM_012475 A150100-USP21_v17 NM_012475_idx1573 113 USP21 NM_012475 A150100-USP21_v2 NM_012475_idx1224 116 USP21 NM_012475 A150100-USP21_v3 NM_012475_idx269 115 USP21 NM_012475 A150100-mmUsp21_v5 NM_013919_idx1120 175 USP21 NM_016572 A150100-USP21_v13 NM_012475_idx741 117 USP21 NM_016572 A150100-USP21_v14 NM_012475_idx928 174 USP21 NM_016572 A150100-USP21_v15 NM_012475_idx682 114 USP21 NM_016572 A150100-USP21_v16 NM_012475_idx733 118 USP21 NM_016572 A150100-USP21_v2 NM_012475_idx1224 116 USP21 NM_016572 A150100-USP21_v3 NM_012475_idx269 115 USP21 NM_016572 A150100-mmUsp21_v5 NM_013919_idx1120 175
 It should be understood that the TARGET genes represented in Table 1 encode different kinds of polypeptides. For example, the TARGETS represented by SEQ ID NO: 40, 43-45 as disclosed herein (Table 1) are GPCRs. Each of these GPCRs is capable of activating an effector protein, resulting in changes in second messenger levels in the cell. The activity of a GPCR can be measured by measuring the activity level of such second messengers. Two important and useful second messengers in the cell are cyclic AMP (cAMP) and Ca2+. The activity levels can be measured by methods known to persons skilled in the art, either directly by ELISA or radioactive technologies or by using substrates that generate a fluorescent or luminescent signal when contacted with Ca2+ or indirectly by reporter gene analysis.
 The activity level of the one or more secondary messengers may typically be determined with a reporter gene controlled by a promoter, wherein the promoter is responsive to the second messenger. Promoters known and used in the art for such purposes are the cyclic-AMP responsive promoter that is responsive for the cyclic-AMP levels in the cell, and the NF-AT responsive promoter that is sensitive to cytoplasmic Ca2+-levels in the cell. The reporter gene typically has a gene product that is easily detectable. The reporter gene can either be stably infected or transiently transfected in the host cell. Useful reporter genes are alkaline phosphatase, enhanced green fluorescent protein, destabilized green fluorescent protein, luciferase and -galactosidase.
 Many of the TARGETS as disclosed herein are kinases and phosphatases, such as the targets represented by SEQ ID NO: 27-34. Specific methods to determine the activity of a kinase or phosphatase by measuring the phosphorylation of a substrate by the kinase or phosphatase, which measurements are performed in the presence or absence of a compound, are well known in the art, whereas some are described in the examples.
 The TARGETS represented by SEQ ID NO: 37, 41, 42, 48, 49, and 51 are proteases. Specific methods to determine the inhibition by the compound by measuring the cleavage of the substrate by the polypeptide, which is a protease, are well known in the art.
 It should be understood that the cells expressing the polypeptides, may be cells naturally expressing the polypeptides, or the cells may be may be transfected to express the polypeptides, as described above.
 In one embodiment it is preferred that the methods of the present invention further comprise the step of contacting the population of cells with an agonist of the polypeptide. This is useful in methods wherein the expression of the polypeptide in a certain chosen population of cells is too low for a proper detection of its activity. By using an agonist the polypeptide may be triggered, enabling a proper read-out if the compound inhibits the polypeptide. Similar considerations apply to the measurement of ECM degradation. In a preferred embodiment, the cells used in the present method are mammalian synovial fibroblasts and the triggers that may be used to induce the ECM-degrading activity are cytokines relevant in the field of arthritis: for instance TNFalpha, IL1beta, IL6, OSM, IL17, and MIF1alpha. In another preferred embodiment, the trigger is a mixture of factors generated by contacting cytokine-producing cells relevant in the field of arthritis, such as monocytes, macrophages, T-cells, and B-cells. The cytokine-producing cells will respond to the contact by producing a complex and unbiased mixture of factors. If the cytokine-producing cell used is also found in a pannus, and the cytokine applied to this trigger is found in the synovial fluid of rheumatoid arthritis patients, the mixture of factors ultimately produced will contain part of the factors that are present in the joints of arthritis patients.
 The present invention further relates to a method for identifying a compound that inhibits extra-cellular matrix degradation, comprising:  (a) contacting a compound with a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 27-51 and 232-295;  (b) determining the binding affinity of the compound to the polypeptide;  (c) contacting a population of mammalian cells expressing said polypeptide with the compound that exhibits a binding affinity of at least 10 micromolar; and  (d) identifying the compound that inhibits extra-cellular matrix degradation.
 The population of cells may be exposed to the compound or the mixture of compounds through different means, for instance by direct incubation in the medium, or by nucleic acid transfer into the cells. Such transfer may be achieved by a wide variety of means, for instance by direct transfection of naked isolated DNA, or RNA, or by means of delivery systems, such as recombinant vectors. Other delivery means such as liposomes, or other lipid-based vectors may also be used. Preferably, the nucleic acid compound is delivered by means of a (recombinant) vector such as a recombinant virus.
 For high-throughput purposes, libraries of compounds may be used such as antibody fragment libraries, peptide phage display libraries, peptide libraries (e.g. LOPAP®, Sigma Aldrich), lipid libraries (BioMol), synthetic compound libraries (e.g. LOPAC®, Sigma Aldrich) or natural compound libraries (Specs, TimTec).
 Preferred drug candidate compounds are low molecular weight compounds. Low molecular weight compounds, i.e. with a molecular weight of 500 Dalton or less, are likely to have good absorption and permeation in biological systems and are consequently more likely to be successful drug candidates than compounds with a molecular weight above 500 Dalton (Lipinski et al. (1997)). Peptides comprise another preferred class of drug candidate compounds. Many GPCRs have a peptide as an agonist or antagonist. Peptides may be excellent drug candidates and there are multiple examples of commercially valuable peptides such as fertility hormones and platelet aggregation inhibitors. Natural compounds are another preferred class of drug candidate compound. Such compounds are found in and extracted from natural sources, and which may thereafter be synthesized. The lipids are another preferred class of drug candidate compound. Many GPCRs have lipids as a ligand.
 Another preferred class of drug candidate compounds is an antibody. The present invention also provides antibodies directed against a TARGET. These antibodies may be endogenously produced to bind to the TARGET within the cell, or added to the tissue to bind to TARGET polypeptide present outside the cell. These antibodies may be monoclonal antibodies or polyclonal antibodies. The present invention includes chimeric, single chain, and humanized antibodies, as well as FAb fragments and the products of a FAb expression library, and Fv fragments and the products of an Fv expression library. In another embodiment, the compound may be a nanobody, the smallest functional fragment of naturally occurring single-domain antibodies (Cortez-Retamozo et al. 2004).
 In certain embodiments, polyclonal antibodies may be used in the practice of the invention. The skilled artisan knows methods of preparing polyclonal antibodies. Polyclonal antibodies can be raised in a mammal, for example, by one or more injections of an immunizing agent and, if desired, an adjuvant. Typically, the immunizing agent and/or adjuvant will be injected in the mammal by multiple subcutaneous or intraperitoneal injections. Antibodies may also be generated against the intact TARGET protein or polypeptide, or against a fragment, derivatives including conjugates, or other epitope of the TARGET protein or polypeptide, such as the TARGET embedded in a cellular membrane, or a library of antibody variable regions, such as a phage display library.
 It may be useful to conjugate the immunizing agent to a protein known to be immunogenic in the mammal being immunized. Examples of such immunogenic proteins include but are not limited to keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. Examples of adjuvants that may be employed include Freund's complete adjuvant and MPL-TDM adjuvant (monophosphoryl Lipid A, synthetic trehalose dicorynomycolate). One skilled in the art without undue experimentation may select the immunization protocol.
 In some embodiments, the antibodies may be monoclonal antibodies. Monoclonal antibodies may be prepared using methods known in the art. The monoclonal antibodies of the present invention may be "humanized" to prevent the host from mounting an immune response to the antibodies. A "humanized antibody" is one in which the complementarity determining regions (CDRs) and/or other portions of the light and/or heavy variable domain framework are derived from a non-human immunoglobulin, but the remaining portions of the molecule are derived from one or more human immunoglobulins. Humanized antibodies also include antibodies characterized by a humanized heavy chain associated with a donor or acceptor unmodified light chain or a chimeric light chain, or vice versa. The humanization of antibodies may be accomplished by methods known in the art (see, e.g. Mark and Padlan, (1994) "Chapter 4. Humanization of Monoclonal Antibodies", The Handbook of Experimental Pharmacology Vol. 113, Springer-Verlag, New York). Transgenic animals may be used to express humanized antibodies.
 Human antibodies can also be produced using various techniques known in the art, including phage display libraries (Hoogenboom and Winter, (1991) J. Mol. Biol. 227:381-8; Marks et al. (1991). J. Mol. Biol. 222:581-97). The techniques of Cole, et al. and Boerner, et al. are also available for the preparation of human monoclonal antibodies (Cole, et al. (1985) Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77; Boerner, et al (1991). J. Immunol., 147(1):86-95).
 Techniques known in the art for the production of single chain antibodies can be adapted to produce single chain antibodies to the TARGET polypeptides and proteins of the present invention. The antibodies may be monovalent antibodies. Methods for preparing monovalent antibodies are well known in the art. For example, one method involves recombinant expression of immunoglobulin light chain and modified heavy chain. The heavy chain is truncated generally at any point in the Fc region so as to prevent heavy chain cross-linking. Alternatively; the relevant cysteine residues are substituted with another amino acid residue or are deleted so as to prevent cross-linking.
 Bispecific antibodies are monoclonal, preferably human or humanized, antibodies that have binding specificities for at least two different antigens and preferably for a cell-surface protein or receptor or receptor subunit. In the present case, one of the binding specificities is for one domain of the TARGET; the other one is for another domain of the same or different TARGET.
 Methods for making bispecific antibodies are known in the art. Traditionally, the recombinant production of bispecific antibodies is based on the co-expression of two immunoglobulin heavy-chain/light-chain pairs, where the two heavy chains have different specificities (Milstein and Cuello, (1983) Nature 305:537-9). Because of the random assortment of immunoglobulin heavy and light chains, these hybridomas (quadromas) produce a potential mixture of ten different antibody molecules, of which only one has the correct bispecific structure. Affinity chromatography steps usually accomplish the purification of the correct molecule. Similar procedures are disclosed in Trauneeker, et al. (1991) EMBO J. 10:3655-9.
 According to another preferred embodiment, the assay method uses a drug candidate compound identified as having a binding affinity for a TARGET, and/or has already been identified as having down-regulating activity such as antagonist activity vis-a-vis one or more TARGET.
 The present invention further relates to a method for inhibiting extra-cellular matrix degradation comprising contacting mammalian cells with an expression inhibitory agent comprising a polyribonucleotide sequence that complements at least about 17 to about 30 contiguous nucleotides of the nucleotide sequence selected from the group consisting of SEQ ID NO: 1-25.
 Another aspect of the present invention relates to a method for inhibiting extra-cellular matrix degradation, comprising by contacting mammalian cells with an expression-inhibiting agent that inhibits the translation in the cell of a polyribonucleotide encoding a TARGET polypeptide. A particular embodiment relates to a composition comprising a polynucleotide including at least one antisense strand that functions to pair the agent with the TARGET mRNA, and thereby down-regulate or block the expression of TARGET polypeptide. The inhibitory agent preferably comprises antisense polynucleotide, a ribozyme, and a small interfering RNA (siRNA), wherein said agent comprises a nucleic acid sequence complementary to, or engineered from, a naturally-occurring polynucleotide sequence selected from the group consisting of SEQ ID NO: 1-25.
 A special embodiment of the present invention relates to a method wherein the expression-inhibiting agent is selected from the group consisting of antisense RNA, antisense oligodeoxynucleotide (ODN), a ribozyme that cleaves the polyribonucleotide coding for SEQ ID NO: 1-25, a small interfering RNA (siRNA, preferably shRNA) that is sufficiently homologous to a portion of the polyribonucleotide corresponding to SEQ ID NO: 1-25, such that the siRNA, preferably shRNA, interferes with the translation of the TARGET polyribonucleotide to the TARGET polypeptide.
 Another embodiment of the present invention relates to a method wherein the expression-inhibiting agent is a nucleic acid expressing the antisense RNA, antisense oligodeoxynucleotide (ODN), a ribozyme that cleaves the polyribonucleotide encoded by SEQ ID NO: 1-25, a small interfering RNA (siRNA, preferably shRNA) that is sufficiently complementary to a portion of the polyribonucleotide corresponding to SEQ ID NO: 1-25, such that the siRNA, preferably shRNA, interferes with the translation of the TARGET polyribonucleotide to the TARGET polypeptide. Preferably the expression-inhibiting agent is an antisense RNA, ribozyme, antisense oligodeoxynucleotide, or siRNA, preferably shRNA, comprising a polyribonucleotide sequence that complements at least about 17 to about 30 contiguous nucleotides of a nucleotide sequence selected from the group consisting of SEQ ID NO: 1-25. More preferably, the expression-inhibiting agent is an antisense RNA, ribozyme, antisense oligodeoxynucleotide, or siRNA, preferably shRNA, comprising a polyribonucleotide sequence that complements at least about 17 to about 25 contiguous nucleotides of a nucleotide sequence selected from the group consisting of SEQ ID NO: 1-25. A special embodiment comprises a polyribonucleotide sequence that complements a polynucleotide sequence selected from the group consisting of SEQ ID NO: 52-175.
 The down regulation of gene expression using antisense nucleic acids can be achieved at the translational or transcriptional level. Antisense nucleic acids of the invention are preferably nucleic acid fragments capable of specifically hybridizing with all or part of a nucleic acid encoding a TARGET polypeptide or the corresponding messenger RNA. In addition, antisense nucleic acids may be designed which decrease expression of the nucleic acid sequence capable of encoding a TARGET polypeptide by inhibiting splicing of its primary transcript. Any length of antisense sequence is suitable for practice of the invention so long as it is capable of down-regulating or blocking expression of a nucleic acid coding for a TARGET. Preferably, the antisense sequence is at least about 17 nucleotides in length. The preparation and use of antisense nucleic acids, DNA encoding antisense RNAs and the use of oligo and genetic antisense is known in the art.
 One embodiment of expression-inhibitory agent is a nucleic acid that is antisense to a nucleic acid comprising SEQ ID NO: 1-25. For example, an antisense nucleic acid (e.g. DNA) may be introduced into cells in vitro, or administered to a subject in vivo, as gene therapy to inhibit cellular expression of nucleic acids comprising SEQ ID NO: 1-25. Antisense oligonucleotides preferably comprise a sequence containing from about 17 to about 100 nucleotides and more preferably the antisense oligonucleotides comprise from about 18 to about 30 nucleotides. Antisense nucleic acids may be prepared from about 17 to about 30 contiguous nucleotides selected from the sequences of SEQ ID NO: 1-25, expressed in the opposite orientation.
 The antisense nucleic acids are preferably oligonucleotides and may consist entirely of deoxyribo-nucleotides, modified deoxyribonucleotides, or some combination of both. The antisense nucleic acids can be synthetic oligonucleotides. The oligonucleotides may be chemically modified, if desired, to improve stability and/or selectivity. Since oligonucleotides are susceptible to degradation by intracellular nucleases, the modifications can include, for example, the use of a sulfur group to replace the free oxygen of the phosphodiester bond. This modification is called a phosphorothioate linkage. Phosphorothioate antisense oligonucleotides are water soluble, polyanionic, and resistant to endogenous nucleases. In addition, when a phosphorothioate antisense oligonucleotide hybridizes to its TARGET site, the RNA-DNA duplex activates the endogenous enzyme ribonuclease (RNase) H, which cleaves the mRNA component of the hybrid molecule.
 In addition, antisense oligonucleotides with phosphoramidite and polyamide (peptide) linkages can be synthesized. These molecules should be very resistant to nuclease degradation. Furthermore, chemical groups can be added to the 2' carbon of the sugar moiety and the 5 carbon (C-5) of pyrimidines to enhance stability and facilitate the binding of the antisense oligonucleotide to its TARGET site. Modifications may include 2'-deoxy, O-pentoxy, O-propoxy, O-methoxy, fluoro, methoxyethoxy phosphorothioates, modified bases, as well as other modifications known to those of skill in the art.
 Another type of expression-inhibitory agent that reduces the levels of TARGETS is the ribozyme. Ribozymes are catalytic RNA molecules (RNA enzymes) that have separate catalytic and substrate binding domains. The substrate binding sequence combines by nucleotide complementarity and, possibly, non-hydrogen bond interactions with its TARGET sequence. The catalytic portion cleaves the TARGET RNA at a specific site. The substrate domain of a ribozyme can be engineered to direct it to a specified mRNA sequence. The ribozyme recognizes and then binds a TARGET mRNA through complementary base pairing. Once it is bound to the correct TARGET site, the ribozyme acts enzymatically to cut the TARGET mRNA. Cleavage of the mRNA by a ribozyme destroys its ability to direct synthesis of the corresponding polypeptide. Once the ribozyme has cleaved its TARGET sequence, it is released and can repeatedly bind and cleave at other mRNAs.
 Ribozyme forms include a hammerhead motif, a hairpin motif, a hepatitis delta virus, group I intron or RNaseP RNA (in association with an RNA guide sequence) motif or Neurospora VS RNA motif. Ribozymes possessing a hammerhead or hairpin structure are readily prepared since these catalytic RNA molecules can be expressed within cells from eukaryotic promoters (Chen, et al. (1992) Nucleic Acids Res. 20:4581-9). A ribozyme of the present invention can be expressed in eukaryotic cells from the appropriate DNA vector. If desired, the activity of the ribozyme may be augmented by its release from the primary transcript by a second ribozyme (Ventura, et al. (1993) Nucleic Acids Res. 21:3249-55).
 Ribozymes may be chemically synthesized by combining an oligodeoxyribonucleotide with a ribozyme catalytic domain (20 nucleotides) flanked by sequences that hybridize to the TARGET mRNA after transcription. The oligodeoxyribonucleotide is amplified by using the substrate binding sequences as primers. The amplification product is cloned into a eukaryotic expression vector.
 Ribozymes are expressed from transcription units inserted into DNA, RNA, or viral vectors. Transcription of the ribozyme sequences are driven from a promoter for eukaryotic RNA polymerase I (pol (I), RNA polymerase II (pol II), or RNA polymerase III (pol III). Transcripts from pol II or pol III promoters will be expressed at high levels in all cells; the levels of a given pol II promoter in a given cell type will depend on nearby gene regulatory sequences. Prokaryotic RNA polymerase promoters are also used, providing that the prokaryotic RNA polymerase enzyme is expressed in the appropriate cells (Gao and Huang, (1993) Nucleic Acids Res. 21:2867-72). It has been demonstrated that ribozymes expressed from these promoters can function in mammalian cells (Kashani-Sabet, et al. (1992) Antisense Res. Dev. 2:3-15).
 A particularly preferred inhibitory agent is a small interfering RNA (siRNA, preferably small hairpin RNA, "shRNA"). siRNA, preferably shRNA, mediate the post-transcriptional process of gene silencing by double stranded RNA (dsRNA) that is homologous in sequence to the silenced RNA. siRNA according to the present invention comprises a sense strand of 17-25 nucleotides complementary or homologous to a contiguous 17-25 nucleotide sequence selected from the group of sequences described in SEQ ID NO: 1-25, preferably from the group of sequences described in SEQ ID No: 52-175, and an antisense strand of 17-25 nucleotides complementary to the sense strand. The most preferred siRNA comprises sense and anti-sense strands that are 100 percent complementary to each other and the TARGET polynucleotide sequence. Preferably the siRNA further comprises a loop region linking the sense and the antisense strand.
 A self-complementing single stranded shRNA molecule polynucleotide according to the present invention comprises a sense portion and an antisense portion connected by a loop region linker. Preferably, the loop region sequence is 4-30 nucleotides long, more preferably 5-15 nucleotides long and most preferably 8 nucleotides long. In a most preferred embodiment the linker sequence is UUGCUAUA (SEQ ID NO: 26; see FIG. 16). Self-complementary single stranded siRNAs form hairpin loops and are more stable than ordinary dsRNA. In addition, they are more easily produced from vectors.
 Analogous to antisense RNA, the siRNA can be modified to confirm resistance to nucleolytic degradation, or to enhance activity, or to enhance cellular distribution, or to enhance cellular uptake, such modifications may consist of modified internucleoside linkages, modified nucleic acid bases, modified sugars and/or chemical linkage the siRNA to one or more moieties or conjugates. The nucleotide sequences are selected according to siRNA designing rules that give an improved reduction of the TARGET sequences compared to nucleotide sequences that do not comply with these siRNA designing rules (For a discussion of these rules and examples of the preparation of siRNA, WO2004094636, published Nov. 4, 2004, and UA20030198627, are hereby incorporated by reference).
 The present invention also relates to compositions, and methods using said compositions, comprising a DNA expression vector capable of expressing a polynucleotide capable of inhibiting extra-cellular matrix degradation and described hereinabove as an expression inhibition agent.
 A special aspect of these compositions and methods relates to the down-regulation or blocking of the expression of a TARGET polypeptide by the induced expression of a polynucleotide encoding an intracellular binding protein that is capable of selectively interacting with the TARGET polypeptide. An intracellular binding protein includes any protein capable of selectively interacting, or binding, with the polypeptide in the cell in which it is expressed and neutralizing the function of the polypeptide. Preferably, the intracellular binding protein is a neutralizing antibody or a fragment of a neutralizing antibody having binding affinity to an epitope of the TARGET polypeptide of SEQ ID NO: 27-51, preferably to a domain of SEQ ID NO: 232-295. More preferably, the intracellular binding protein is a single chain antibody.
 A special embodiment of this composition comprises the expression-inhibiting agent selected from the group consisting of antisense RNA, antisense oligodeoxynucleotide (ODN), a ribozyme that cleaves the polyribonucleotide coding for SEQ ID NO: 27-51, and a small interfering RNA (siRNA) that is sufficiently homologous to a portion of the polyribonucleotide corresponding to SEQ ID NO: 1-25, such that the siRNA interferes with the translation of the TARGET polyribonucleotide to the TARGET polypeptide,
 The polynucleotide expressing the expression-inhibiting agent is preferably included within a vector. The polynucleic acid is operably linked to signals enabling expression of the nucleic acid sequence and is introduced into a cell utilizing, preferably, recombinant vector constructs, which will express the antisense nucleic acid once the vector is introduced into the cell. A variety of viral-based systems are available, including adenoviral, retroviral, adeno-associated viral, lentiviral, herpes simplex viral or a sendaviral vector systems, and all may be used to introduce and express polynucleotide sequence for the expression-inhibiting agents in TARGET cells.
 Preferably, the viral vectors used in the methods of the present invention are replication defective. Such replication defective vectors will usually pack at least one region that is necessary for the replication of the virus in the infected cell. These regions can either be eliminated (in whole or in part), or be rendered non-functional by any technique known to a person skilled in the art. These techniques include the total removal, substitution, partial deletion or addition of one or more bases to an essential (for replication) region. Such techniques may be performed in vitro (on the isolated DNA) or in situ, using the techniques of genetic manipulation or by treatment with mutagenic agents. Preferably, the replication defective virus retains the sequences of its genome, which are necessary for encapsidating, the viral particles.
 In a preferred embodiment, the viral element is derived from an adenovirus. Preferably, the vehicle includes an adenoviral vector packaged into an adenoviral capsid, or a functional part, derivative, and/or analogue thereof. Adenovirus biology is also comparatively well known on the molecular level. Many tools for adenoviral vectors have been and continue to be developed, thus making an adenoviral capsid a preferred vehicle for incorporating in a library of the invention. An adenovirus is capable of infecting a wide variety of cells. However, different adenoviral serotypes have different preferences for cells. To combine and widen the TARGET cell population that an adenoviral capsid of the invention can enter in a preferred embodiment, the vehicle includes adenoviral fiber proteins from at least two adenoviruses. Preferred adenoviral fiber protein sequences are serotype 17, 45 and 51. Techniques or construction and expression of these chimeric vectors are disclosed in US Published Patent Applications 20030180258 and 20040071660, hereby incorporated by reference.
 In a preferred embodiment, the nucleic acid derived from an adenovirus includes the nucleic acid encoding an adenoviral late protein or a functional part, derivative, and/or analogue thereof. An adenoviral late protein, for instance an adenoviral fiber protein, may be favorably used to TARGET the vehicle to a certain cell or to induce enhanced delivery of the vehicle to the cell. Preferably, the nucleic acid derived from an adenovirus encodes for essentially all adenoviral late proteins, enabling the formation of entire adenoviral capsids or functional parts, analogues, and/or derivatives thereof. Preferably, the nucleic acid derived from an adenovirus includes the nucleic acid encoding adenovirus E2A or a functional part, derivative, and/or analogue thereof. Preferably, the nucleic acid derived from an adenovirus includes the nucleic acid encoding at least one E4-region protein or a functional part, derivative, and/or analogue thereof, which facilitates, at least in part, replication of an adenoviral derived nucleic acid in a cell. The adenoviral vectors used in the examples of this application are exemplary of the vectors useful in the present method of treatment invention.
 Certain embodiments of the present invention use retroviral vector systems. Retroviruses are integrating viruses that infect dividing cells, and their construction is known in the art. Retroviral vectors can be constructed from different types of retrovirus, such as, MoMuLV ("murine Moloney leukemia virus" MSV ("murine Moloney sarcoma virus"), HaSV ("Harvey sarcoma virus"); SNV ("spleen necrosis virus"); RSV ("Rous sarcoma virus") and Friend virus. Lentiviral vector systems may also be used in the practice of the present invention. Retroviral systems and herpes virus system may be preferred vehicles for transfection of neuronal cells.
 In other embodiments of the present invention, adeno-associated viruses ("AAV") are utilized. The AAV viruses are DNA viruses of relatively small size that integrate, in a stable and site-specific manner, into the genome of the infected cells. They are able to infect a wide spectrum of cells without inducing any effects on cellular growth, morphology or differentiation, and they do not appear to be involved in human pathologies.
 In the vector construction, the polynucleotide agents of the present invention may be linked to one or more regulatory regions. Selection of the appropriate regulatory region or regions is a routine matter, within the level of ordinary skill in the art. Regulatory regions include promoters, and may include enhancers, suppressors, etc.
 Promoters that may be used in the expression vectors of the present invention include both constitutive promoters and regulated (inducible) promoters. The promoters may be prokaryotic or eukaryotic depending on the host. Among the prokaryotic (including bacteriophage) promoters useful for practice of this invention are lac, lacZ, T3, T7, lambda Pr, P1, and trp promoters. Among the eukaryotic (including viral) promoters useful for practice of this invention are ubiquitous promoters (e.g. HPRT, vimentin, actin, tubulin), intermediate filament promoters (e.g. desmin, neurofilaments, keratin, GFAP), therapeutic gene promoters (e.g. MDR type, CFTR, factor VIII), tissue-specific promoters (e.g. actin promoter in smooth muscle cells, or Flt and Flk promoters active in endothelial cells), including animal transcriptional control regions, which exhibit tissue specificity and have been utilized in transgenic animals: elastase I gene control region which is active in pancreatic acinar cells (Swift, et al. (1984) Cell 38:639-46; Ornitz, et al. (1986) Cold Spring Harbor Symp. Quant. Biol. 50:399-409; MacDonald, (1987) Hepatology 7:425-515); insulin gene control region which is active in pancreatic beta cells (Hanahan, (1985) Nature 315:115-22), immunoglobulin gene control region which is active in lymphoid cells (Grosschedl, et al. (1984) Cell 38:647-58; Adames, et al. (1985) Nature 318:533-8; Alexander, et al. (1987) Mol. Cell. Biol. 7:1436-44), mouse mammary tumor virus control region which is active in testicular, breast, lymphoid and mast cells (Leder, et al. (1986) Cell 45:485-95), albumin gene control region which is active in liver (Pinkert, et al. (1987) Genes and Devel. 1:268-76), alpha-fetoprotein gene control region which is active in liver (Krumlauf, et al. (1985) Mol. Cell. Biol., 5:1639-48; Hammer, et al. (1987) Science 235:53-8), alpha 1-antitrypsin gene control region which is active in the liver (Kelsey, et al. (1987) Genes and Devel., 1: 161-71), beta-globin gene control region which is active in myeloid cells (Mogram, et al. (1985) Nature 315:338-40; Kollias, et al. (1986) Cell 46:89-94), myelin basic protein gene control region which is active in oligodendrocyte cells in the brain (Readhead, et al. (1987) Cell 48:703-12), myosin light chain-2 gene control region which is active in skeletal muscle (Sani, (1985) Nature 314.283-6), and gonadotropic releasing hormone gene control region which is active in the hypothalamus (Mason, et al. (1986) Science 234:1372-8).
 Other promoters which may be used in the practice of the invention include promoters which are preferentially activated in dividing cells, promoters which respond to a stimulus (e.g. steroid hormone receptor, retinoic acid receptor), tetracycline-regulated transcriptional modulators, cytomegalovirus immediate-early, retroviral LTR, metallothionein, SV-40, E1a, and MLP promoters.
 Additional vector systems include the non-viral systems that facilitate introduction of polynucleotide agents into a patient. For example, a DNA vector encoding a desired sequence can be introduced in vivo by lipofection. Synthetic cationic lipids designed to limit the difficulties encountered with liposome-mediated transfection can be used to prepare liposomes for in vivo transfection of a gene encoding a marker (Felgner, et. al. (1987) Proc. Natl. Acad. Sci. USA 84:7413-7); see Mackey, et al. (1988) Proc. Natl. Acad. Sci. USA 85:8027-31; Ulmer, et al. (1993) Science 259:1745-8). The use of cationic lipids may promote encapsulation of negatively charged nucleic acids, and also promote fusion with negatively charged cell membranes (Feigner and Ringold, (1989) Nature 337:387-8). Particularly useful lipid compounds and compositions for transfer of nucleic acids are described in International Patent Publications WO 95/18863 and WO 96/17823, and in U.S. Pat. No. 5,459,127. The use of lipofection to introduce exogenous genes into the specific organs in vivo has certain practical advantages and directing transfection to particular cell types would be particularly advantageous in a tissue with cellular heterogeneity, for example, pancreas, liver, kidney, and the brain. Lipids may be chemically coupled to other molecules for the purpose of TARGETing. Targeted peptides, e.g., hormones or neurotransmitters, and proteins for example, antibodies, or non-peptide molecules could be coupled to liposomes chemically. Other molecules are also useful for facilitating transfection of a nucleic acid in vivo, for example, a cationic oligopeptide (e.g., International Patent Publication WO 95/21931), peptides derived from DNA binding proteins (e.g., International Patent Publication WO 96/25508), or a cationic polymer (e.g., International Patent Publication WO 95/21931).
 It is also possible to introduce a DNA vector in vivo as a naked DNA plasmid (see U.S. Pat. Nos. 5,693,622, 5,589,466 and 5,580,859). Naked DNA vectors for therapeutic purposes can be introduced into the desired host cells by methods known in the art, e.g., transfection, electroporation, microinjection, transduction, cell fusion, DEAE dextran, calcium phosphate precipitation, use of a gene gun, or use of a DNA vector transporter (see, e.g., Wilson, et al. (1992) J. Biol. Chem. 267:963-7; Wu and Wu, (1988) J. Biol. Chem. 263:14621-4; Hartmut, et al. Canadian Patent Application No. 2,012,311, filed Mar. 15, 1990; Williams, et al (1991). Proc. Natl. Acad. Sci. USA 88:2726-30). Receptor-mediated DNA delivery approaches can also be used (Curiel, et al. (1992) Hum. Gene Ther. 3:147-54; Wu and Wu, (1987) J. Biol. Chem. 262:4429-32).
 The present invention also provides biologically compatible, extra-cellular matrix degradation inhibiting compositions comprising an effective amount of one or more compounds identified as TARGET inhibitors, and/or the expression-inhibiting agents as described hereinabove.
 A biologically compatible composition is a composition, that may be solid, liquid, gel, or other form, in which the compound, polynucleotide, vector, and antibody of the invention is maintained in an active form, e.g., in a form able to effect a biological activity. For example, a compound of the invention would have inverse agonist or antagonist activity on the TARGET; a nucleic acid would be able to replicate, translate a message, or hybridize to a complementary mRNA of a TARGET; a vector would be able to transfect a TARGET cell and expression the antisense, antibody, ribozyme or siRNA as described hereinabove; an antibody would bind a TARGET polypeptide domain.
 A preferred biologically compatible composition is an aqueous solution that is buffered using, e.g., Tris, phosphate, or HEPES buffer, containing salt ions. Usually the concentration of salt ions will be similar to physiological levels. Biologically compatible solutions may include stabilizing agents and preservatives. In a more preferred embodiment, the biocompatible composition is a pharmaceutically acceptable composition. Such compositions can be formulated for administration by topical, oral, parenteral, intranasal, subcutaneous, and intraocular, routes. Parenteral administration is meant to include intravenous injection, intramuscular injection, intraarterial injection or infusion techniques. The composition may be administered parenterally in dosage unit formulations containing standard, well-known non-toxic physiologically acceptable carriers, adjuvants and vehicles as desired.
 A particularly preferred embodiment of the present composition invention is a extra-cellular matrix degradation inhibiting pharmaceutical composition comprising a therapeutically effective amount of an expression-inhibiting agent as described hereinabove, in admixture with a pharmaceutically acceptable carrier. Another preferred embodiment is a pharmaceutical composition for the treatment or prevention of a condition involving ECM degradation, or a susceptibility to the condition, comprising an effective extra-cellular matrix degradation inhibiting amount of a TARGET antagonist or inverse agonist, its pharmaceutically acceptable salts, hydrates, solvates, or prodrugs thereof in admixture with a pharmaceutically acceptable carrier.
 Pharmaceutical compositions for oral administration can be formulated using pharmaceutically acceptable carriers well known in the art in dosages suitable for oral administration. Such carriers enable the pharmaceutical compositions to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, and the like, for ingestion by the patient. Pharmaceutical compositions for oral use can be prepared by combining active compounds with solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are carbohydrate or protein fillers, such as sugars, including lactose, sucrose, mannitol, or sorbitol; starch from corn, wheat, rice, potato, or other plants; cellulose, such as methyl cellulose, hydroxypropylmethyl-cellulose, or sodium carboxymethyl-cellulose; gums including arabic and tragacanth; and proteins such as gelatin and collagen. If desired, disintegrating or solubilizing agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, alginic acid, or a salt thereof, such as sodium alginate. Dragee cores may be used in conjunction with suitable coatings, such as concentrated sugar solutions, which may also contain gum arabic, talc, polyvinyl-pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for product identification or to characterize the quantity of active compound, i.e., dosage.
 Pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a coating, such as glycerol or sorbitol. Push-fit capsules can contain active ingredients mixed with filler or binders, such as lactose or starches, lubricants, such as talc or magnesium stearate, and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid, or liquid polyethylene glycol with or without stabilizers.
 Preferred sterile injectable preparations can be a solution or suspension in a non-toxic parenterally acceptable solvent or diluent. Examples of pharmaceutically acceptable carriers are saline, buffered saline, isotonic saline (e.g. monosodium or disodium phosphate, sodium, potassium; calcium or magnesium chloride, or mixtures of such salts), Ringer's solution, dextrose, water, sterile water, glycerol, ethanol, and combinations thereof 1,3-butanediol and sterile fixed oils are conveniently employed as solvents or suspending media. Any bland fixed oil can be employed including synthetic mono- or di-glycerides. Fatty acids such as oleic acid also find use in the preparation of injectables.
 The composition medium can also be a hydrogel, which is prepared from any biocompatible or non-cytotoxic homo- or hetero-polymer, such as a hydrophilic polyacrylic acid polymer that can act as a drug absorbing sponge. Certain of them, such as, in particular, those obtained from ethylene and/or propylene oxide are commercially available. A hydrogel can be deposited directly onto the surface of the tissue to be treated, for example during surgical intervention.
 Embodiments of pharmaceutical compositions of the present invention comprise a replication defective recombinant viral vector encoding the polynucleotide inhibitory agent of the present invention and a transfection enhancer, such as poloxamer. An example of a poloxamer is Poloxamer 407, which is commercially available (BASF, Parsippany, N.J.) and is a non-toxic, biocompatible polyol. A poloxamer impregnated with recombinant viruses may be deposited directly on the surface of the tissue to be treated, for example during a surgical intervention. Poloxamer possesses essentially the same advantages as hydrogel while having a lower viscosity.
 The active expression-inhibiting agents may also be entrapped in microcapsules prepared, for example, by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsules and poly-(methylmethacylate) microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences (1980) 16th edition, Osol, A. Ed.
 Sustained-release preparations may be prepared. Suitable examples of sustained-release preparations include semi-permeable matrices of solid hydrophobic polymers containing the antibody, which matrices are in the form of shaped articles, e.g. films, or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and gamma-ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOT®. (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(-)-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods. When encapsulated antibodies remain in the body for a long time, they may denature or aggregate as a result of exposure to moisture at 37° C., resulting in a loss of biological activity and possible changes in immunogenicity. Rational strategies can be devised for stabilization depending on the mechanism involved. For example, if the aggregation mechanism is discovered to be intermolecular S--S bond formation through thio-disulfide interchange, stabilization may be achieved by modifying sulfhydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions.
 As defined above, therapeutically effective dose means that amount of protein, polynucleotide, peptide, or its antibodies, agonists or antagonists, which ameliorate the symptoms or condition. Therapeutic efficacy and toxicity of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., ED50 (the dose therapeutically effective in 50% of the population) and LD50 (the dose lethal to 50% of the population). The dose ratio of toxic to therapeutic effects is the therapeutic index, and it can be expressed as the ratio, LD50/ED50. Pharmaceutical compositions that exhibit large therapeutic indices are preferred. The data obtained from cell culture assays and animal studies is used in formulating a range of dosage for human use. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage varies within this range depending upon the dosage form employed, sensitivity of the patient, and the route of administration.
 For any compound, the therapeutically effective dose can be estimated initially either in cell culture assays or in animal models, usually mice, rabbits, dogs, or pigs. The animal model is also used to achieve a desirable concentration range and route of administration. Such information can then be used to determine useful doses and routes for administration in humans. The exact dosage is chosen by the individual physician in view of the patient to be treated. Dosage and administration are adjusted to provide sufficient levels of the active moiety or to maintain the desired effect. Additional factors which may be taken into account include the severity of the disease state, age, weight and gender of the patient; diet, desired duration of treatment, method of administration, time and frequency of administration, drug combination(s), reaction sensitivities, and tolerance/response to therapy. Long acting pharmaceutical compositions might be administered every 3 to 4 days, every week, or once every two weeks depending on half-life and clearance rate of the particular formulation.
 The pharmaceutical compositions according to this invention may be administered to a subject by a variety of methods. They may be added directly to targeted tissues, complexed with cationic lipids, packaged within liposomes, or delivered to targeted cells by other methods known in the art. Localized administration to the desired tissues may be done by direct injection, transdermal absorption, catheter, infusion pump or stent. The DNA, DNA/vehicle complexes, or the recombinant virus particles are locally administered to the site of treatment. Alternative routes of delivery include, but are not limited to, intravenous injection, intramuscular injection, subcutaneous injection, aerosol inhalation, oral (tablet or pill form), topical, systemic, ocular, intraperitoneal and/or intrathecal delivery. Examples of ribozyme delivery and administration are provided in Sullivan et al. WO 94/02595.
 Antibodies according to the invention may be delivered as a bolus only, infused over time or both administered as a bolus and infused over time. Those skilled in the art may employ different formulations for polynucleotides than for proteins. Similarly, delivery of polynucleotides or polypeptides will be specific to particular cells, conditions, locations, etc.
 As discussed hereinabove, recombinant viruses may be used to introduce DNA encoding polynucleotide agents useful in the present invention. Recombinant viruses according to the invention are generally formulated and administered in the form of doses of between about 104 and about 1014 pfu. In the case of AAVs and adenoviruses, doses of from about 106 to about 1011 pfu are preferably used. The term pfu ("plaque-forming unit") corresponds to the infective power of a suspension of virions and is determined by infecting an appropriate cell culture and measuring the number of plaques formed. The techniques for determining the pfu titre of a viral solution are well documented in the prior art.
 The present invention also provides methods of inhibiting extra-cellular matrix degradation, comprising administering, to a subject suffering from a disease condition involving extra-cellular matrix degradation, an extra-cellular matrix degradation inhibiting pharmaceutical composition as described herein, preferably a therapeutically effective amount of an expression-inhibiting agent of the present invention. The diseases involving extra-cellular marix degradation, include psoriatic arthritis, juvenile arthritis, early arthritis, reactive arthritis, osteoarthritis, ankylosing spondylitis. osteoporosis, muskulo skeletal diseases such as tendinitis and periodontal disease, cancer metastasis, airway diseases (COPD, asthma), renal and liver fibrosis, cardio-vascular diseases such as atherosclerosis and heart failure, and neurological diseases such as neuroinflammation and multiple sclerosis. More preferred diseases for treatment in accordance with the present invention are the degenerative joint diseases such as psoriatic arthritis, juvenile arthritis, early arthritis, reactive arthritis, osteoarthritis, ankylosing spondylitis. The most preferred degenerative joint disease for treatment in accordance with the present method is rheumatoid arthritis,
 Administering of the expression-inhibiting agent of the present invention to the subject patient includes both self-administration and administration by another person. The patient may be in need of treatment for an existing disease or medical condition, or may desire prophylactic treatment to prevent or reduce the risk for diseases and medical conditions affected by a disturbance in bone metabolism. The expression-inhibiting agent of the present invention may be delivered to the subject patient orally, transdermally, via inhalation, injection, nasally, rectally or via a sustained release formulation.
 A preferred regimen of the present method comprises the administration to a subject in suffering from a disease condition characterized by inflammatory, with an effective inhibiting amount of an expression-inhibiting agent of the present invention for a period of time sufficient to reduce the abnormal levels of extracellular matrix degradation in the patient, and preferably terminate, the self-perpetuating processes responsible for said degradation. A special embodiment of the method comprises administering of an effective matrix metallo-protease inhibiting amount of a expression-inhibiting agent of the present invention to a subject patient suffering from or susceptible to the development of rheumatoid arthritis, for a period of time sufficient to reduce or prevent, respectively, collagen and bone degradation in the joints of said patient, and preferably terminate, the self-perpetuating processes responsible for said degradation.
 The invention also relates to the use of an agent as described above for the preparation of a medicament for treating or preventing a disease involving extra-cellular matrix degradation.
 Preferably the pathological condition is arthritis. More preferably, the pathological condition is rheumatoid arthritis.
 The polypeptides and polynucleotides useful in the practice of the present invention described herein may be free in solution, affixed to a solid support, borne on a cell surface, or located intracellularly. To perform the methods it is feasible to immobilize either the TARGET polypeptide or the compound to facilitate separation of complexes from uncomplexed forms of the polypeptide, as well as to accommodate automation of the assay. Interaction (e.g., binding of) of the TARGET polypeptide with a compound can be accomplished in any vessel suitable for containing the reactants. Examples of such vessels include microtitre plates, test tubes, and microcentrifuge tubes. In one embodiment, a fusion protein can be provided which adds a domain that allows the polypeptide to be bound to a matrix. For example, the TARGET polypeptide can be "His" tagged, and subsequently adsorbed onto Ni-NTA microtitre plates, or ProtA fusions with the TARGET polypeptides can be adsorbed to IgG, which are then combined with the cell lysates (e.g., (35)s-labelled) and the candidate compound, and the mixture incubated under conditions favorable for complex formation (e.g., at physiological conditions for salt and pH). Following incubation, the plates are washed to remove any unbound label, and the matrix is immobilized. The amount of radioactivity can be determined directly, or in the supernatant after dissociation of the complexes. Alternatively, the complexes can be dissociated from the matrix, separated by SDS-PAGE, and the level of the protein binding to the TARGET protein quantified from the gel using standard electrophoretic techniques.
 Other techniques for immobilizing protein on matrices can also be used in the method of identifying compounds. For example, either the TARGET or the compound can be immobilized utilizing conjugation of biotin and streptavidin. Biotinylated TARGET protein molecules can be prepared from biotin-NHS (N-hydroxy-succinimide) using techniques well known in the art (e.g., biotinylation kit, Pierce Chemicals, Rockford, Ill.), and immobilized in the wells of streptavidin-coated 96 well plates (Pierce Chemical). Alternatively, antibodies reactive with the TARGETS but which do not interfere with binding of the TARGET to the compound can be derivatized to the wells of the plate, and the TARGET can be trapped in the wells by antibody conjugation. As described above, preparations of a labeled candidate compound are incubated in the wells of the plate presenting the TARGETS, and the amount of complex trapped in the well can be quantitated.
 The polynucleotides encoding the TARGET polypeptides are identified as SEQ ID NO: 1-25. Applicants have shown that transfection of mammalian cells with these polynucleotides in an expressible form increase the release of factors that promote extra-cellular matrix degradation.
 The present invention also relates to a method for diagnosis of a pathological condition involving ECM degradation, comprising determining the nucleic acid sequence of at least one of the genes of SEQ ID NO: 1-25 within the genomic DNA of a subject; comparing the sequence with the nucleic acid sequence obtained from a database and/or a healthy subject; and identifying any difference(s) related to the onset of the pathological condition.
 Still another aspect of the invention relates to a method for diagnosing a pathological condition involving extra-cellular matrix degradation or a susceptibility to the condition in a subject, comprising determining the amount of polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 27-51 in a biological sample, and comparing the amount with the amount of the polypeptide in a healthy subject, wherein an increase of the amount of polypeptide compared to the healthy subject is indicative of the presence of the pathological condition.
 The invention is further illustrated in the following figures and examples.
 The following assays, when used in combination with arrayed adenoviral libraries (the production and use of which are described in WO99/64582), are useful for the discovery of factors that modulate the capacity of synovial fibroblasts (SFs) to degrade collagen, the main component of cartilage. Candidate factors are filtered first through a primary followed by a secondary assay. Example 1 describes the development and setup of the primary assay screen of an adenoviral cDNA library using an ELISA for detection of protein levels of Matrix Metalloprotease 1 (MMP1), and is referred to herein as the "MMP1 assay". Example 2 describes the screening and its results. Examples 3 and 4 describe the secondary assay, which is more functionally oriented, detects collagen degradation in the supernatant of SFs, and is referred to herein as the "collagen degradation assay". Example 5 describes the testing for the endogenous expression of factors in SFs. This method in referred to as "expression profiling" of hits in various RA-derived SFs (RASFs). Example 6 describes the effect of the reduction in activity of various genes on the cytokine-induced SF MMP1 expression thereby determining the collagenolytic activity of RASF's.
Control Viruses Used:
 The control viruses used in these studies are listed below. dE1/dE2A adenoviruses are generated from these adapter plasmids by co-transfection of the helper plasmid pWEAd5AflII-rITR.dE2A in PER.E2A packaging cells, as described in WO99/64582.
(A) Negative Control Viruses:
 Ad5-LacZ: Described as pIPspAdApt6-lacZ in WO02/070744.  Ad5-ALPP: The 1.9 kb insert is isolated from pGT65-PLAP (Invitrogen) by digestion with NsiI; blunted; followed by digestion with EcoRI and cloned into EcoRI and HpaI-digested pIPspAdApt6.  Ad5-eGFP: Described as pIPspAdApt6-EGFP in WO02/070744.
TABLE-US-00004  Ad5-eGFP_KD: Target sequence: GCTGACCCTGAAGTTCATC. (SEQ ID NO: 179)
 Cloned using Sap1-sites into vector and virus generated as described in WO03/020931.
TABLE-US-00005  Ad5-Luciferase_KD_v13: Target sequence: GCTGACCCTGAAGTTCATC. (SEQ ID NO: 180)
Cloned using Sap1-sites into vector and virus generated as described in WO03/020931.
TABLE-US-00006 Ad5-M6PR_KD_v1: Target sequence: GCTGACCCTGAAGTTCATC. (SEQ. ID NO: 296)
Cloned using Sap1-sites into vector and virus generated as described in WO03/020931.
(B) Positive Control Viruses:
 Ad5-RELA: The cDNA encoding RELA is obtained by PCR on a human placenta cDNA library with the following primers:
TABLE-US-00007  upstream: GCGAAGCTTGCGGCATGGACGAACTGT (SEQ ID NO: 181) and downstream: GCGGGATCCCAGGCGTCACCCCCTTAG. (SEQ ID NO: 182)
 A 1681 bp DNA insert is generated of which the 5' sequence corresponds to NM--021975. Primers are designed such that the PCR products can be inserted into the pIPspAdapt6 vector by HindIII-BamHI cloning.
 Ad5-MMP1: The cDNA encoding MMP1, cloned into the pIPspAdapt6 plasmid, is isolated from a human placenta cDNA library (see WO02/070744) by classical filter colony hybridisation strategy. A human placental cDNA library is transformed into bacteria and plated out on agar plates. Thousands of individual colonies are picked (using a Q-pix device (Genetix)) and re-arrayed on agar plates. After growing bacteria up, these plates are overlayed on hybridisation filters. These filters are subjected to a classical hybridisation procedure with a MMP1 specific probe. This probe is obtained by PCR on a placenta cDNA library using the following primers:
TABLE-US-00008 upstream: GTTCTGGGGTGTGGTGTCTCACAGC; (SEQ ID NO: 183) and downstream: CAAACTGAGCCACATCAGGCACTCC. (SEQ ID NO: 184)
 A bacterial colony, at a position corresponding to that of a positive signal spot on the filter after hybridisation, is picked and used for plasmid preparation. 5' sequence verification confirms that the 5' sequence of the insert corresponds to NM--002421.  Ad5-TRAF6: The cDNA encoding TRAF6 is isolated according to the same colony hybridisation technique as the one described for MMP1. The TRAF6 specific probe is obtained by PCR on a placenta cDNA library using the following primers:
TABLE-US-00009  upstream: CCAGTCTGAAAGTGACTGCTGTGTGG; (SEQ ID NO: 185) and downstream: CAACTGGACATTTGTGACCTGCATCC. (SEQ ID NO: 186)
 A bacterial colony, at a position corresponding to that of a positive signal spot on the filter after hybridisation, is picked and used for plasmid preparation. 5' sequence verification confirms that the 5' sequence of the insert corresponds to NM--004620.2.  Ad5-MMP13: The cDNA of MMP13 is isolated from a cDNA preparation from human synovial fibroblasts by PCR. The 1498 by PCR product is cloned into pIPspAdapt6 using a HindIII/EcoRI cloning strategy. Sequence verification confirms that the insert corresponds to by 18 to 1497 of NM--002427.
 Ad5-MYD88: This cDNA is isolated from a human placenta cDNA library constructed in pIPspAdapt6. The virus mediating the expression of MYD88 is identified as a hit in one of the genomic screen run at Galapagos Genomics. Sequence verification of the insert confirms that the insert corresponds to by 40 to 930 of NM--002468.
 Ad5TNFRIA: This virus is isolated from a human placenta cDNA library constructed in pIPspAdapt6. The virus mediating the expression of MYD88 is identified as a hit in one of the genomic screen run at Galapagos Genomics. 5' sequence verification of the 1.4 Kb insert reveals that the insert starts at by 958 of NM--001065. Virus is generated as described in WO03/020931.
TABLE-US-00010 Ad5-MMP1_KD_v10: Target sequence: GCTGACCCTGAAGTTCATC. (SEQ ID NO: 187)
Cloned using Sap1-sites into vector and virus generated as described in WO03/020931.
Development of the MMP Assay
 Matrix Metallo Proteases (MMPs) possess various physiological roles, for example, they are involved in the maturation of other proteases, growth factors, and the degradation of extra-cellular matrix components. MMP1 is a member of the MMP family and is able to degrade native collagen, the main component of bone and cartilage. Increased expression of MMP1 by synovial fibroblasts (SFs) is diagnostic for the progression of the arthritic disease and is predictive for erosive processes in the joint (Cunnane et al., 2001). SF expression of MMP1 can be increased by the activation of SFs with triggers relevant for rheumatoid arthritis, such as the cytokines TNF-α and IL1β (Andreakos et al., 2003). The measurement of the MMP1 levels produced by activated SFs is highly relevant in the context of RA as this event reflects the level of activation of SFs towards an erosive phenotype as it is seen in the pannus. If reduced expression of a candidate target protein in activated SFs leads to the reduction of MMP1 expression in these cells, then the target is shown to be involved in the regulation of MMP1 expression and thus considered relevant for the development of therapeutic strategies for the treatment of RA. The identification of such target proteins involves the screening of a collection of recombinant adenoviruses mediating the expression of a library of cDNAs, further referred to as "Ad-cDNAs". The collection used herein is further referred to as "adenoviral cDNA library" or the "FlexSelect collection" (see WO99/64582).
 The MMP1 assay is developed by first testing the capacity of Synovial fibroblasts (SFs) to produce MMP1.
 A. To evaluate the capacity of SFs to produce MMP1, a set of adenoviruses mediating the expression of TRAF6 and MYD88, adaptor molecules in the IL1 pathway, and p65/RelA, a subunit of the NF B transcription factor that is known to increase expression of factors involved in the immune and inflammatory responses, both of which are expected to increase MMP1 expression (see Vincenti and Brinckerhoff, 2002) are used to infect SFs.
 40,000 SFs are seeded per well of a 6-well plate in DMEM+10% FBS and infected with a multiplicity of infection (MOI) of 7500 viral particles per cell (vp/cell). The expression of MMP1 by SFs is first determined at the mRNA level, by means of real-time, quantitative PCR. RNA of the cells infected with the control viruses is prepared 48 h post infection using the SV RNA isolation kit (Promega), according to the instructions of the manufacturer. cDNA is prepared from this RNA using Multiscribe reverse transcriptase (50 U/μl, Applied Biosystems) and random hexamers. cDNA synthesis is performed in 25 μl total volume consisting of 1×TaqMan buffer A (PE Applied Biosystems), 5 mM MgCl2, 500 mM total dNTPs, 2.5 mM random hexamers, 0.4 U/μl RNase Inhibitor, and 1.25 U/μl MultiScribe Reverse Transcriptase. The mixture is incubated for 10 min at 25° C., 30 min at 48° C., and 5 min at 95° C. Specific DNA products are amplified from the resulting cDNA with AmpliTaq Gold DNA polymerase (Applied BioSystems) during 40 PCR cycles using suited primer pairs. Amplification of the specific DNA products is monitored on an ABI PRISM® 7000 Sequence Detection System. The subsequent real time PCR reaction contained 5 μl of the RT reaction product in a total volume of 25 μl consisting of 1×SYBR Green mix (Applied Biosystems), 300 nM forward primer, and 300 nM reverse primer. Each sample is analyzed in duplicate. The PCR reaction is performed using the following program: 10 min at 95° C. followed by 40 cycles of (15 sec 95° C., 1 min 60° C.). After each PCR reaction the products are analysed by measuring the dissociation curve by incubating for 15 sec 95° C., and 15 sec at 60° C., followed by increasing the temperature to 95° C. over a 20 min time period, ending with 15 sec at 95° C. The sequences of the primer pairs used for the detection of MMP1, 18S and -actin expression are listed in Table 2.
TABLE-US-00011 TABLE 2 List of primers and their sequences used herein. SEQ Hit Primer ID number name Primer sequence NO NA pAdapt_FW GGTGGGAGGTCTATATAAGC 188 pAdapt_REV GGACAAACCACAACTAGAATGC 189 NA MMP2_For CCCCAGGCACTGGTGTTG 190 MMP2_Rev ACGGACCACTTGGCCTTCT 191 NA MMP1_For CCGGTTTTTCAAAGGGAATAAGTAC 192 MMP1_Rev TTCACAGTTCTAGGGAAGCCAAAG 193 H31-031 CAMK4_For CAGCATCCGTGGGTCACA 194 CAMK4_Rev TTCACCGCTGCCTTAAGCTT 195 H31-035 PRKCE_For TGAGGACGACCTATTTGAGTCCAT 196 PRKCE_Rev GGGATTCTTCGTCATGAAAGCT 197 H31-047 USP21_For CTGCGAAGCTGTGAATCCTACTC 198 USP21_Rev GGCATCCTGCTGGCTGTATC 199 H31-049 CASP10_For TCCTGGCAGAACTCCTCTATATCATAC 200 CASP10_Rev TGACAGTTCGTAGAGCAGGTTTCTA 201 H31-180 TM7SF1_For GAACTTGTACTTCACGCAGGTG 202 TM7SF1_Rev CAACAGGAAAACAAGGCTGATG 203 H31-242 GPR21_For TGCGTGGTCCCTTCTTTATCAC 204 GPR21_Rev GCCATGGAGACGCTCTTCAG 205 H31-290 RIPK2_For CATTAAATGAACTCCTACATAGGAAAAC 206 RIPK2_Rev AGGGCAATTTCATGCAGGAT 207 H31-301 TPST1_For GGAGTGTCTCTGTCAAAAGTGGA 208 TPST1_Rev ACCCATTTTGATAGAGCTCCTACATT 209 H31-319 MST3_For GACATTAAAGCGGCCAACGT 210 MST3_Rev CTCGGGTGCCATCCAGAA 211 H31-347 SEPT1_For GCGAGAAAGACGAAGAGCTGC 212 SEPT1_Rev GCCTGGCTCTGCTGCATT 213 H31-351 CD72_For CAGTGAAATTTATCCACAATCACAC 214 CD72_Rev AGAGCTGAGGCCAGTCCAATAT 215 H31-360 RIT_For GGTGTAGGGAAGAGTGCCATGA 216 RIT_Rev GCATCTTCAATGGTGGGATCA 217 H31-384 FXYD5_For TGGTCGCCTGTGTCTTCTCA 218 FXYD5_Rev GTGGTATCTTTCAACGTCTGTCCTC 219 H31-450 Q9ESW8_For GAGGAAGGCGGTGGTAGTGA 220 Q9ESW8_Rev CTCAACCGGAATCTCGTACACA 221 H34-067 FZD6_For TGGGAGATAACTTGGGTCTCTGAT 222 FZD6_Rev AAGCCAATTCTGGTCGAGCTT 223 H34-087 MKNK1_For AGGGAGCCTATGCCAAAGTTC 224 MKNK1_Rev CTCGATGATTTTGACGGCATAC 225 H34-088 MAPKAPK5-- GAGGAAGCTCCTGAAGGTCAAAC 226 For MAPKAPK5-- CAACCACTGCCTTGTCCATC 227 Rev H34-092 FZD4_For AGCCAGCTGCAGTTCTTCCTT 228 FZD4_Rev TCACAGCGTCTCTTGACTGAAAG 229
 MMP1 is detected using the SYBR Green method, whereas the levels of 18S rRNA, used as internal calibrator for the PCR reaction, is measured using a Taqman probe (TaqMan® Ribosomal RNA Control Reagents, Applied Biosystems). The amplification plot and the resulting threshold Ct value are indicators for the amount of specific mRNAs present in the samples. Delta-delta Ct values are presented, meaning the normalized (relative to the 18S calibrator) levels of MMP1 mRNA in the samples infected with the positive control viruses relative to the expression levels in a Ad5-eGFP infected control sample. Results indicate a strong up-regulation of the MMP1 mRNA levels upon expression of p65/RelA, TRAF6 or MYD88 in SFs as compared to the non-infected or Ad5-eGFP-infected SFs.
 The level of MMP1 expressed by SFs is also determined at the protein level by Western Blotting. Two days after infection, supernatant of cells, infected with various recombinant adenoviruses as indicated for the Real-time PCR experiment, is collected and concentrated 15 times by classical TCA precipitation. 15 μl of the supernatant are resolved by SDS-PAGE using a 10% polyacrylamide gel. For these experiments, the medium used is M199 medium+1% FBS. For the MMP1 control sample, non-concentrated supernatant of cells infected with Ad5-MMP1 is loaded onto the gel. The resolved proteins are transferred onto a nitrocellulose membrane. The quality of the transfer and equal loading of the samples are verified by Ponceau-S staining of the membrane. Immunodetection is performed using a goat anti-MMP1 polyclonal antibody as primary antibody (R&D Systems, 1/500 dilution) and an HRP-linked rabbit anti-goat antibody (DAKO, 1/10000 dilution) as secondary antibody and ECL plus HRP substrate (Amersham Biosciences). The Western Blotting revealed a strongly increased level of MMP1 protein in the supernatant of the SFs infected with the adenoviruses mediating expression of Ad5-p65/RelA, Ad5-TRAF6 or Ad5-MYD88 as compared to the Ad5-eGFP infected cells. A very strong signal is detected for the supernatant of cells infected with Ad5-MMP1 (FIG. 2, panels B and C).
 The high levels of MMP1 protein present in the supernatant of the Ad5-p65/RelA, Ad5-TRAF6 or Ad5-MYD88 infected SFs are confirmed using a commercially available MMP1 activity ELISA (RPN2629, Amersham Biosciences). In this ELISA, MMP1 is captured by an antibody immobilized in a well and the amount is subsequently quantified based on the conversion of a MMP1 substrate. 50 μl of non-concentrated supernatant of SFs (prepared as indicated for the western blotting experiment) are processed in this ELISA as recommended by the manufacturer.
 These experiments confirm the capacity of SFs, in general, and of the cell batch used for screening and validation experiments, to produce MMP1 protein upon triggering of inflammatory pathways.
 A 384-well format ELISA for measurement of MMP1 is developed. Various primary antibodies are tested, as well as various ELISA protocols. The following protocol is developed and validated to measure MMP1 levels in SF supernatant in 384 well plates: white Lumitrac 600 384 well plates (Greiner) are coated with 2 μg/ml anti-MMP1 antibody MAB1346 (Chemicon). The antibody is diluted in buffer 40 (1.21 g Tris base (Sigma), 0.58 g NaCl (Calbiochem) and 5 ml 10% NaN3 (Sigma) in 1 L milliQ water and adjusted to pH 8.5). After overnight incubation at 4° C., plates are washed with PBS (80 g NaCl, 2 g KCl (Sigma), 11.5 g Na2HPO4.7H2O and 2 g KH2PO4 in 10 L milliQ; pH 7.4) and blocked with 100 μl/well Casein buffer (2% Casein (VWR International) in PBS). Next day, casein buffer is removed from ELISA plates and replaced by 50 μl/well EC buffer (4 g casein, 2.13 g Na2HPO4 (Sigma), 2 g bovine albumin (Sigma), 0.69 g NaH2PO4.H2O (Sigma), 0.5 g CHAPS (Roche), 23.3 g NaCl, 4 ml 0.5 M EDTA pH 8 (Invitrogen), 5 ml 10% NaN3 in 1 L milliQ and adjusted to pH 7.0). 0.25 mM DTT (Sigma) is added to the thawed samples plates. After removal of the EC buffer, 20 μl of sample is transferred to the ELISA plates. After overnight incubation at 4° C. plates are washed twice with PBS and once with PBST (PBS with 0.05% Tween-20 (Sigma)) and incubated with 35 μl/well biotinylated anti-MMP1 antibody solution (R&D). This secondary antibody is diluted in buffer C (0.82 g NaH2PO4.H2O, 4.82 g Na2HPO4, 46.6 g NaCl, 20 g bovine albumin and 4 ml 0.5M EDTA pH 8 in 2 L milliQ and adjusted to pH 7.0) at a concentration of 5 μg/ml. After 2 h of incubation at RT, plates are washed as described above and incubated with 50 μl/well streptavidin-HRP conjugate (Biosource). Streptavidin-HRP conjugate is diluted in buffer C at a concentration of 0.25 μg/ml. After 45 min, plates are washed as described above and incubated for 5 min with 50 μl/well BM Chem ELISA Substrate (Roche). Readout is performed on the Luminoscan Ascent Luminometer (Labsystems) with an integration time of 200 msec or with an Envision reader (Perkin Elmer).
 Typical results obtained with the MMP1 ELISA developed are shown in FIG. 3. For this experiment, 3000 SFs are seeded in a 96 well plate in DMEM+10% FBS. 24 h later, SFs are either infected at an MOI of 10000 with adenoviruses mediating the expression of ALPP, MYD88, MMP1; or left uninfected. One day after the infection, the medium of the cells is replaced by M199 medium (Invitrogen) supplemented with 1% FBS. After an incubation time of 48 hrs, the supernatant is harvested, transferred to a 384 well plate and subjected to the MMP1 ELISA procedure described above. A robust, more than 3.5-fold up-regulation of the signal is observed. This experiment demonstrated the robustness and specificity of the MMP1 ELISA.
 The increase of MMP1 expression by SFs upon treatment with cytokines relevant in the field of RA (TNF, IL1 and OSM) or a combination thereof is monitored. Results are shown in FIG. 10 as white bars. For this experiment, SFs are seeded in 96 well plates at 3000 cells/well. 24 h later, the medium is changed to M199 medium supplemented with 1% FBS. One day after the medium change, cytokines or combinations thereof are added to the cultures, each cytokine being added to a final concentration of 25 ng/ml. 72 h after cytokine addition, the supernatant is collected and processed in the ELISA, as described for FIG. 3. As shown in FIG. 10, white bars, TNF alone induces an almost 3-fold increase in MMP1 expression. Triggering of SFs with a combination of TNF and OSM and/or IL1 leads to even higher MMP1 expression levels. This experiment demonstrates that the sensitivity of the MMP1 ELISA developed is sufficient to measure increases in MMP1 expression by SFs driven by cytokines involved in RA pathogenesis.
Screening of 4224 Recombinant Adenoviruses in an MMP1 Assay
 A 384 well control plate is generated to assess the quality of the assay during the different screening runs. The composition of this plate is shown in FIG. 4A. Wells are filled with control viruses that are produced under the same conditions as the FlexSelect adenoviral cDNA library. This control plate contains three sets of 48 positive control viruses (P1 (Ad5-MMP1), P2 (Ad5-TRAF6), P3 (Ad5-MYD88)), arranged in diagonal, interspaced with three sets of 48 negative control viruses (N1 (Ad5-eGFP), N2 (Ad5-LacZ), N3 (Ad5-ALPP), Bl: blanco, uninfected). Every well contains 50 μl of virus crude lysate. The viruses contained in the control plate are generated according to the protocol applied for the construction of the FlexSelect collection. Multiple aliquots of this control plate are produced and stored at -80° C.
 Optimal screening protocol: RASFs are cultured in DMEM medium (Invitrogen) supplemented with 10% fetal calf serum (ICN), 100 units/ml penicillin (Invitrogen) and 100 μg/ml streptomycin (Invitrogen) and incubated at 37° C. and 10% CO2. The cells are passed once a week by a 1/3 split. The maximal passage number for RASFs used in the screening is 11. For screening, SFs are seeded in transparent 384 well plates (Greiner) coated with 0.1% gelatin (Merck) at a density of 1500 cells/well in 25 μl Synovial Cell growth medium (Cell Applications, Inc.). After overnight incubation, cells are infected with 3 μl Ad-cDNA from the Galapagos FlexSelect adenoviral cDNA library. As the average titer of the adenoviral library is 3×109 virus particles/ml, this represents an MOI of 6000. 24 h after infection, the medium is changed to 50 μl of M199 medium supplemented with 1% FCS. 40 μl supernatant is collected 72 h later into new transparent 384 well plates (Greiner) and stored at -80° C. until further processing in the MMP1 ELISA. The infection, medium change and medium collection steps are performed with a TECAN Freedom pipettor. The ELISA step is performed as indicated in Example 1.
 A representative example of the performance of the control plate tested with the protocol described above is shown in FIG. 4B. Synovial fibroblasts are infected with 3 μl of the viruses contained in the control plate in an arrayed fashion using a TECAN 384 channel pipettor. The medium is refreshed the day after infection and the supernatant is harvested after 72 h production time and subjected to the 384 well format MMP1 ELISA described in previous example. The raw luminescence signal obtained is shown.
 A stringent cutoff is applied, that is the average of all 144 negative control viruses plus 4.5 times the standard deviation over these samples. As expected, the Ad5-MMP1 control virus scored very well in the assay, with all 48 Ad5-MMP1 viruses being picked up as a hit above this cutoff. The Ad5-MYD88 control virus also scored robustly, with 84% of the Ad5-MYD88 control viruses being picked up above the applied cutoff. The weaker Ad5-TRAF6 control, which gave rise to weaker increases in MMP1 mRNA levels (see Example 1) did not perform strongly, indicating that this cutoff will likely identify strong MMP1 inducers.
 The MMP1 assay on RASFs described above is screened against the adenoviral cDNA libraries (FlexSelect® collection) developed at Galapagos Genomics. The main part of this adenoviral collection contains cDNAs of genes from "drugable" classes like GPCRs, kinases, proteases, phosphodiesterases and nuclear hormone receptors. The majority of these cDNAs are obtained by a PCR-based approach briefly described below. Based on the sequences of the selected genes, which are obtained from the RefSeq database, PCR primers are designed for amplification of the complete open reading frame from ATG start codon to the stop codon. Primers are received in an arrayed format with forward and reverse primers mixed at a PCR ready concentration in 96 well plates. From this point on, the arrayed format is maintained throughout all the handlings (from PCR till virus production) resulting in an arrayed adenoviral cDNA library. As a template for the PCR reactions, placental, fetal liver, fetal brain and spinal cord cDNA libraries are used (from Invitrogen or Edge Biosystems). For the genes encoded by a single exon, PCR reactions are performed on human genomic DNA. After the amplification reactions, the size of the PCR products is estimated and compared to the predicted size based on sequence information. The PCR products obtained are purified with a 96-well PCR clean-up system (Wizard magnesil, Promega, Madison, Wis., USA), digested with the appropriate restriction enzymes (AscI, NotI or SalI restriction sites are included in the primers) and directly cloned into the adenoviral adapter plasmid pIspAdAdapt-10-Zeo (described in U.S. Pat. No. 6,340,595) using DNA ligation kit version 2 (TaKaRa, Berkeley, Calif., USA). After a transformation and selection step, multiple clones per gene, one of which is sequence verified, are used for the preparation of plasmid DNA and subsequent generation of adenovirus according to the procedure described in WO99/64582.
 The total FlexSelect adenoviral cDNA library consisted of 11×384 well plates at the time it is screened. 4224 samples represents 1705 genes.
 The MMP1 assay is screened against the FlexSelect adenoviral cDNA library using the optimized protocol described above. Every cDNA library plate is screened in duplicate in a primary screen and in a rescreen. As such, four data points are obtained for each cDNA clone. A representative example of screening results and of the analysis performed to identify hits is shown in FIG. 5.
 SFs are seeded in 384 well plates and infected with 3 μl of 384 different recombinant adenoviruses of the FlexSelect collection contained in an arrayed fashion (using a TECAN pipetor), in a 384 well plate. The medium is refreshed the day after infection; the supernatant is harvested after 72 h production time and subjected to the MMP1 ELISA using a luminescent substrate. The raw luminescence signal obtained is shown. For every individual virus, the viruses mediating the expression of PRKCE, CASP10 and USP21 in particular, the 2 datapoints (FIGS. 5A and B) obtained in the primary screen (FIG. 5A) and in the rescreen (FIG. 5B) are shown.
 To determine the cutoff value for hit calling, the average as well as standard deviation are calculated on all data points obtained per screening batch after removal of the 10% highest and 10% lowest values. The cutoff value is then defined as 3 times the standard deviation added to the average. This cutoff is indicated as a horizontal line in the graph in FIG. 5. Screening and rescreening results are presented in FIG. 6 for 4 cDNA encoding PRKCE, 5 cDNAs encoding USP21 and 4 cDNAs encoding CASP10. All 4 PRKCE cDNA clones scored above cutoff in duplicate in both the primary screen and rescreen, 4 out of 5 USP21 clones scored above cutoff in primary screening and rescreening, and 3 out of 4 CASP10 cDNA clones scored in duplicate in primary screening and rescreening. These data are indicative of the quality of the screening and of the FlexSelect cDNA collection.
 As mentioned, every screening plate is screened and rescreened in duplicate. Only samples that scored above the cutoff value (the average plus 3 times standard deviation) for 3 out of the 4 datapoints are selected as hits. In addition, if multiple clones scored positive, maximally 2 clones per gene are further processed through the collagen degradation assay. As such, 253 hit Ad-cDNAs, representing 229 genes, are finally picked, propagated and tested in the collagen degradation assay.
 `Knock-in viruses` mediating the expression of various target genes listed in Table 1 are tested as follows. On day 1, SFs are seeded, in Synovial growth medium, in gelatin coated 96 well plates at a density of 3000 cells per well or in 384 well plates at a density of 1500 cells per well. 1 day after seeding, the cells are infected at the volumes or MOIs indicated on the figures. On day 3, the medium is refreshed to M199 medium supplemented with 1% FBS. On day 6, the supernatant is collected and subjected to the MMP1 ELISA according to the protocol described above. The Ad5-Luciferase, Ad5-eGFP or Ad5-Empty viruses are used as negative control viruses. Infection of SFs with recombinant adenoviruses driving the expression of SEPT1, TPST1, USP21, MKNK1, RIPK2 (FIG. 13 A), PGPEP1, RITZ (FIG. 13 B), CAMK4, MST3, PRKCE (FIG. 13 C) and CD72, TM7SF1, GPR21 (FIG. 13 D) clearly mediated an increased expression of MMP1 by the infected SFs. The results shown in FIG. 13 are the averages of duplicate datapoints.
Development of a Screening Method for the Measurement of the Collagenolytic Activity of Primary Synovial Fibroblasts (SFs): Collagen Degradation Assay
 The MMP1 assay is used as a first filter to select hits that mediated an increase in the MMP1 expression in SFs. The amount of MMP1 present in the supernatant of SFs might not, however, be sufficient to mediate the degradation of native collagen. In addition, besides MMP1, additional proteases might be expressed by SFs that, alone or in synergy with MMP1, mediate collagen breakdown. In order to rank our hits according to their potential to increase the collagenolytic activity of SFs, the present inventors developed a functional assay that determines the extent of degradation of native collagen in the supernatant of SFs. The various reagents and buffers used to perform the assay described below are from Chondrex (Redmond, USA), unless mentioned otherwise.
 In first instance, the assay is developed to be compatible with a cDNA library screening on primary human cells. As a second development step, the assay is miniaturized to be compatible with an arrayed, medium throughput assay. Experiments confirmed that the sensitivity of the collagen assay performed on primary cells in miniaturized configuration is conserved as compared to the assay in non-miniaturized configuration. The results of a typical experiment illustrating this finding are shown in FIG. 6. For this experiment, SFs (seeded at a density of 3000 cells/well in a 96 well plate in M199 medium supplemented with 1% FBS) are infected (MOI 10,000) with Ad5-ALPP, AD5-TRAF6 or Ad5-MYD88. After an incubation time of 48 hrs (post infection), the supernatant is harvested and tested in both the miniaturized and non-miniaturized collagen degradation assays. Fluorescence signal, which is proportional to the level of collagen degradation, is indicated.
 "Non-miniaturized" collagen degradation assay protocol: 100 μl of the SF supernatant or 100 μl of M199 medium+1% FBS supplemented with the indicated amount of rMMP1 (R&D systems) or chymotrypsin (Sigma) are mixed with 90 μl of buffer B. These mixes are added to either 10 μl of trypsin activating solution, or 10 μl of APMA (4-aminophenyl mercuric acetate, 2 mM final, Sigma) activating solution. These activating solutions mediate the removal of the pro-domain of MMPs that keep these proteases in an inactive state. In the case of trypsin activation, the mixture is incubated for 60 min at 35° C., followed by the addition of SBTI (soybean trypsin inhibitor) to inactivate all non-collagenolytic proteases, whereas in the case of APMA activation, the mixture is incubated for 10 min at 35° C. 100 μl of Buffer A and 100 μl of native FITC-labeled bovine collagen type I (1 mg/ml, in 0.01N acetic acid) are mixed and added to the activated samples followed by an incubation step of 2 h at 35° C. during which collagenases cleave the FITC-labeled collagen in the typical 1/4 and 3/4 fragments. The reaction is stopped by addition of 10 μl of the stop solution (1,10 phenantroline, 10 mM final, Sigma). The large collagenase pieces are further digested by the addition of 10 μl of elastase (the "enhancer" solution) and incubation for 30 min at 35° C. After cooling down the samples, 400 μl of extraction buffer is added to precipitate the non-cleaved collagen fragments. These fragments are separated from the digested collagen pieces by a centrifugation step (10,000 rpm, 10 min). 200 μl sample is transferred to a black 96 well plate for a fluorescence measurement (520 nm, 480 nm as emission and excitation wavelengths, respectively) performed on a Fluostar reader (BMG).
 "Miniaturized collagen degradation assay" protocol: A 96 well plate (V-bottom, Greiner) is filled with 9 μl of solution B and 1 μl of trypsin solution per well. 10 μl of sample is added per well, followed by incubation for 15 min at 34° C. After incubation, 1 μl SBTI is added. 20 μl of FITC-Collagen mix (10 μl FITC-labeled collagen type I+10 μl solution A) are added to the activated sample followed by incubation for 24 h at 34° C. One μl of 1.10 Phenantroline (Sigma) is added to the reaction mixture. One μl of enhancer solution (elastase) is added, followed by incubation for 30 min at 34° C. When the reaction mixture is at room temperature, 40 μl extraction buffer are added and the plate is sealed (Nunc seals) and vortexed. After centrifugation for 25 min at 4000 rpm (Beckman centrifuge), 50 μl of the supernatant are transferred into a black F-bottom plate (Greiner) and fluorescence is measured on a Fluostar reader (BMG), 480 nm excitation wavelength, 520 nm emission wavelength). The results of the experiment are shown in FIG. 8, and shows increased collagen type I degradation in the supernatant of Ad5-TRAF6 as well as Ad5-MYD88 infected cells. As such, the 2 positive controls identified for the "MMP1 assay" on SFs also mediate increased collagenolytic capacity of SFs. This suggests that the potency of a cDNA in the "MMP1 assay" is predictive for its capacity to increase the global collagenolytic activity of SFs. Although the levels of the fluorescent signal in the miniaturized assay are lower as compared to the non-miniaturized assay, the relative increase in fluorescence in the positive samples as compared to the Ad5-ALPP control is maintained. Thus, a miniaturized collagen degradation assay on SFs has been developed that has a sensitivity level comparable to the non-miniaturized assay. This result establishes that the method used for the collagen degradation assay described above is compatible with the screening of cDNA libraries (in adenoviral format in this example) on primary cells (human SFs in this example). Various experiments established that following aspects of the protocol are important:  the use of trypsin for the activation of the latent MMPs in the supernatant of the cells is useful for the detection of collagenase activity using the assay.  the supernatant of non-infected cells does not contain any detectable background collagenase activity. It is held that the use of medium without phenol red (M199 medium, no phenol red, Invitrogen) with low serum content (1% FBS) is preferred to obtain this low background signal.  the collagen used for this assay is mostly in native, triple helix conformation, as no collagen degradation is mediated by chymotrypsin, an enzyme that has the capacity to degrade denatured collagen (gelatin). The native character of the collagen used is also preferred for this assay.
 The above miniaturized assay is compared to another low-throughput detection method for collagen degradation, in which the following samples are tested: supernatant of SFs (cultured in 96 well plates in M199 medium supplemented with 1% FBS) uninfected or infected with Ad5-ALPP, Ad5-TRAF6, Ad5-PRKCD, Ad5-MMP13 (MMP13 is a potent collagenase), or Ad5-TNFR1A, all at an MOI of 10,000. The results of the miniaturized collagen degradation assay run on these samples (following the protocol described in former example) is shown in FIG. 7: Supernatant obtained after infection of SFs with the indicated recombinant adenoviruses and a 48 hrs production time, is subjected to both the miniaturized (fluorescence-based) collagen degradation assay and the lower throughput visual assessment of collagen degradation. For the latter test, the various supernatants are incubated with native collagen. The reaction mixtures are resolved on a polyacrylamide gel and degradation of the heterotrimeric collagen type I fibrils from the native (bands A and B) to the 3/4 N-terminal TCA fragments (bands C and D) is assessed after Coomassie staining
 A cutoff value for hits versus non-hits in this experiment is defined as the average over the data points for the uninfected control samples plus 3 times the standard deviation over these data points and is indicated as a dotted line on the bar graph in FIG. 7. These data indicate that, in addition to the Ad5-TRAF6 and Ad5-MMP13 positive controls, the collagenolytic potential of SFs increased upon overexpression of PRKCD and TNFR1A. As TNFalpha is a well-known trigger involved in RA pathogenesis, it can be expected that the overexpression of TNFR1A, the TNFa receptor, will lead to an increase in collagen degradation. This result further validates our approach to identify relevant cDNAs involved in RA pathogenesis. In this experiment, PRKCD is identified as another relevant mediator of collagen degradation by SFs.
 The same samples are then tested in the following setup: a 10 μl sample is mixed with 10 μl EDANS buffer (50 mM Tris-HCl, pH 7.5; 150 mM NaCl; 10 mM CaCL2; 0.05% Brij-35, 50 μM ZnCl2), 10 μl of a solution of collagen type I (IBFB, Germany, 1 mg/ml dissolved in 0.01N acetic acid). APMA is added to this reaction mixture to a final concentration of 2 mM. The reaction mixture is incubated for 48 h at 35° C. 25 μl of the reaction mixture is then boiled and resolved on a 8% SDS poly acryl amide gel (Novex) which is then subjected to a coomassie blue staining Native collagen type I is a triple helix composed of 21 and 12 chains. These chains are visible on the gel in the control samples and are indicated by arrows A and B in the lower part of FIG. 7. In the positive control samples, Ad5-MMP13, Ad5-TRAF6 and Ad5-PRKCD, these 2 bands are cleaved into the 3/4N-terminal "TCA" fragments, indicated by arrows C and D. This typical restriction pattern is indicative for the action of MMP-type collagenases, which cleaves the collagen triple helix at a single position, thereby generating characteristic 1/4 C-terminal "TCB" and 3/4 N-terminal "TCA" fragments. These results confirm in a visual way the direct relationship that exists between the signal obtained in the collagen degradation assay and the collagen degrading activity present in the tested samples. These data also confirm that the signal obtained in the collagen degradation assay is the result of the activity of MMP-type collagenases.
 As the main component of cartilage is collagen type II, we compared the collagen degradation assay readout performed with FITC-labeled collagen type I and with FITC-labeled collagen type II. Results of a representative experiment are shown in FIG. 8. For this experiment, supernatant is used of SFs (cultured in 96 wells plates, 3000 cells/well in M199+1% FBS) that are infected with Ad5-TRAF6, Ad5-ALPP or Ad5-MYD88 at an MOI of 10,000. Supernatant of these cells is harvested 48 h post-infection and subjected to the non-miniaturized collagen degradation assay procedure described for FIG. 8 with either FITC-labeled native collagen type I or FITC-labeled collagen type II (same amounts as FITC-labeled collagen type I). Results shown in FIG. 8 indicate that the degradation of collagen type II gave rise to lower fluorescent signals, suggesting a higher resistance of collagen type II to proteolytic degradation as compared to collagen type I. Notwithstanding the lower signal levels obtained when using collagen type II, cDNAs mediating increased collagen type II degradation are identified, as exemplified here with Ad5-TRAF6. The order of potency of the hits towards induction of collagen degradation is maintained in the collagen degradation assay run with collagen type II as compared to the assay run with collagen type I. These results indicate that the capacity of a hit to induce the degradation of collagen type I in this assay is predictive for its capacity to induce the degradation of collagen type II.
Testing of 253 Hits of the "MMP1 Assay" and Screening of 1679 Recombinant Adenoviruses in the Collagen Degradation Assay
 The adenoviruses identified as hits in the MMP1 assay on primary synovial fibroblasts (SFs) are picked from the FlexSelect adenoviral cDNA library and are re-propagated in 96 well plate format by infection of PER.E2A producer cells (see WO99/64582). These plates are further referred to as the "MMP1 hit propagation plates". On these plates, 4 Ad5-ALPP and 4 Ad5-Luciferase control viruses are also included. The border wells of these plates are not used to avoid eventual "border effects" in the experiments. The MMP1 hit propagation plates contain 50 hit viruses and 10 negative control viruses. This virus material is then tested at 3 MOI's in duplicate in the collagen type I degradation assay on SFs as follows. SFs are trypsinized and seeded in 96 well plates (Nunc, transparent plates, tissue culture treated). Trypsinized SFs are resuspended in Synoviocyte Growth medium (Cell Applications) at a density of 30,000 cells/ml and 100 l of this suspension is dispensed in each well using a multidrop dispenser (Labsystems). Approximately 24 h after seeding of the cells, a duplicate infection of the cells is performed with 6, 12 or 18 μl of the virus material present in 96 well MMP1 hit propagation plates using a Tecan Freedom 200 pipettor (Tecan). As such, the content of the MMP1 hit propagation plates is transferred to 6 96 well plates containing the seeded SFs. 6 data points in the collagen degradation assay are generated per hit virus. Approximately 24 h after infection, virus and medium are removed from the cells using an 8 channel Vacusafe device (Integra) and 60 μl M199 medium supplemented with +0.5% FBS is added to every well.
 72 h after medium refreshment, supernatant is transferred to a 96 well plate (V-bottom, Greiner) with the Tecan Freedom 200 pipettor. The supernatant is stored at -80° C. until use. To perform the assay, the supernatant is thawed and the assay is performed according to the protocol of the miniaturized collagen type I degradation assay described above in Example 3.
 Hit selection is performed as follows: For each plate, the average and standard deviation are calculated for the fluorescence measurements obtained for the 8 wells infected with control viruses. The cutoff for hits versus no-hit is defined as the average plus 2 times the standard deviation for these control samples. A virus is considered a hit if it induced a signal above the cutoff value for at least 3 out of 6 data points. 253 hits identified in the MMP1 assay have been retested according to this procedure. Out of these, 61 Ad-cDNAs significantly increased the collagenolytic activity of SFs, representing 55 individual genes when redundancy is taken into account. Besides these 55 hits, two Ad-cDNAs picked up in the screening delivered a proof of principle for the screening. One of these hits encoded MMP1. Another hit encodes IKK (IKBKB). This kinase has a central role in the response of cells to inflammatory triggers as e.g. TNF. Small drug inhibitors, with RA as therapeutic indication, are currently being designed against IKK (Andreakos et al., 2003). The fact that hits, relevant in the field of RA, are picked up confirms the quality of our screening concept and the quality if the materials (assays and libraries) used.
 As final quality control on these hit Ad-cDNAs, their identity is checked by sequence analysis. The procedure for sequence analysis is as follows. The hit viruses are propagated using PER.E2A producer cells in a 96 well plate. PER.E2A cells are seeded in 96 well plates at a density of 40,000 cells/well in 180 μl medium. Cells are incubated overnight at 39° C. in a 10% CO2 humidified incubator. One day later, cells are infected with 1 μl of crude cell lysate from FlexSelect stocks containing the hit Ad-cDNA. Cells are incubated further at 34° C., 10% CO2 until appearance of cytopathic effect (as revealed by the swelling and rounding up of the cells, typically 7 days post infection). The supernatant is collected and the virus crude lysate is treated with proteinase K: 12 μl crude lysate is added to 4 μl Lysis buffer (1× Expand High Fidelity buffer with MgCl2 (Roche Molecular Biochemicals, Cat. No 1332465) supplemented with 1 mg/ml proteinase K (Roche Molecular Biochemicals, Cat No 745 723) and 0.45% Tween-20 (Roche Molecular Biochemicals, Cat No 1335465) in sterile PCR tubes. These are incubated at 55° C. for 2 h followed by a 15 min inactivation step at 95° C. For the PCR reaction, 1 μl lysate is added to a PCR master mix composed of 5 μl 10× Expand High Fidelity buffer with MgCl2, 0.5 μl of dNTP mix (10 mM for each dNTP), 1 μl of `Forward primer` (10 mM stock, sequence: 5' GGT GGG AGG TCT ATA TAA GC; SEQ ID NO: 230), 1 μl of `Reverse Primer` (10 mM stock, sequence: 5' GGA CAA ACC ACA ACT AGA ATG C; SEQ ID NO: 231), 0.2 μl of Expand High Fidelity DNA polymerase (3.5 U/μl, Roche Molecular Biochemicals) and 41.3 μl of H2O.
 PCR is performed in a PE Biosystems GeneAmp PCR system 9700 as follows: the PCR mixture (50 μl in total) is incubated at 95° C. for 5 min; each cycle runs at 95° C. for 15 sec, 55° C. for 30 sec, 68° C. for 4 min, and is repeated for 35 cycles. A final incubation at 68° C. is performed for 7 min. 5 μl of the PCR mixture is mixed with 2 μl of 6× gel loading buffer, loaded on a 0.8% agarose gel containing 0.5 μg/μl ethidium bromide to resolve the amplification products. The size of the amplified fragments is estimated from a standard DNA ladder loaded on the same gel. For sequencing analysis, the cDNAs expressed by the target adenoviruses are amplified by PCR using primers complementary to vector sequences flanking the SapI site of the pIPspAdapt6 plasmid. The sequence of the PCR fragments is determined and compared with the expected sequence.
Screening of the FlexSelect Collection Subset in the Collagen Degradation Assay
 The possibility exists that certain factors mediate an increased collagenolytic activity of SFs through collagenases other than MMP1. In order to identify such factors, a subset of the FlexSelect collection is screened in the collagen degradation assay on SFs. 384 well plates from the FlexSelect collection containing mainly Ad-cDNAs mediating the expression of kinases and GPCRs are screened. The following screening protocol is applied. SFs are trypsinized and resuspended in Synoviocyte Growth medium (Cell Applications) at a density of 30,000 cells/ml. 100 μl of this cell suspension is dispensed in each well of 96 well plates (Nunc, tissue culture treated) using a `multidrop` dispenser (Labsystems). Approximately 24 h after seeding of the cells, they are infected with the library Ad-cDNAs as follows. The FlexSelect library aliquot plates (384 well format, stored at -80° C.) to be processed are thawed at RT in a laminar air flow cabinet for 1 h. Plates are then stored at 4° C. until further processing.
 For every well of a quadrant of a 384-well adenoviral cDNA library aliquot plate, 10 μl of virus crude lysate is transferred to a well of a 96 well plate containing the SFs. This action is performed with the 96 needle head of a TECAN Freedom 200 pipettor. Each virus is assayed in duplicate. As such, for every 384-well virus library aliquot plates, 8 96-well plates containing SF are infected. In between every pipeting step, needles of the pipettor are emptied in a bleach wash station and rinsed two times with 175 μl of bleach (5%) and two times with 200 μl of water and finally with 200 μl of ethanol (20%). Approximately 24 h after infection, the medium of the cells is refreshed. Virus and medium are removed with the Vacusafe (Integra) and 60 μl of fresh M199 medium+0.5% FBS is added. 72 h after refreshment of the medium, the cell supernatant is transferred from the 96 well plates containing the infected SFs to a 96 well plate (V-bottom, Greiner) with the TECAN Freedom 200 pipettor. The samples are then subjected to the miniaturized collagen type I degradation assay. In total, 1679 samples are screened in duplicate in this assay, representing 449 genes.
 The following analysis is performed for hit selection: Per screening batch, the average and standard deviation is calculated on all samples after removal of the 10% highest and 10% lowest values. As mentioned above, 2 data points are obtained for every Ad-cDNA sample screened. The Ad-cDNA samples for which one of the 2 data points scored above the average plus 4 times the standard deviation as well as the samples for which both data points scored above the average plus 2 times standard deviation are selected as hits. A representative example of the results obtained during screening for 96 viruses (1 assay plate) screened in duplicate is shown in FIG. 9. For every individual virus, the 2 datapoints (A and B) obtained in the primary screen are shown. Viruses mediating the expression of CASP10 and MMP3 are indicated. The signal obtained for the samples is expressed relative to the standard deviation and average using following formula: [Times standard deviation difference from average=(Value Sample-Value Average)/Standard deviation]. The cutoff for hit calling (average plus 2 or 4 times standard deviation) is indicated as a full or dotted line, respectively. Among the 96 Ad-cDNAs for which screening results are shown, 4, out of which 3 scored according to the selection criterion, mediated the expression of MMP3 and 4, out of which 3 scored according to the selection criterion, mediated the expression of CASP10. 108 Ad-cDNAs, representing 79 genes when taking redundancy into account, are selected as hits according to this procedure.
 These hits are re-propagated and rescreened using the procedure described for the screening of the hits of the MMP1 assay in the collagen degradation assay. 31 hits, representing 20 individual genes, out of the 108 primary hits mediated a significant level of collagen type I degradation in the rescreen procedure. As 4 genes out of the 55 identified as hits through the "MMP1 assay" and validated in the collagen degradation assay are also present among the 20 genes identified as hits in the screening of a subset of the FlexSelect collection in the collagen degradation assay, a total of 71 genes are identified that increased the collagenolytic potential when expressed or activated in primary human SFs. The preferred hit genes identified in this assay are listed in Table 1. The performance of these in the collagen degradation assay in summarized in Table 4.
TABLE-US-00012 TABLE 4 Summary of the Features of the TARGET Genes Experiment Description Knock-in Knock down Knock down Knock-in Induction of Expression Inhibition of Inhibition of cytokine Gene MMP1 collagen in primary cytokine induced induced collagen Symbol induction degradation RASFs MMP1 degradation RIPK2 SP SP SP SP NT PRKCE SP SP P SP SP MST3 SP SP P P NT MAPKAPK5 N N P SP SP MKNK1 SP SP P N NT CAMK4 P P P SP SP SEPT1 P P P SP NT PGPEP1 P P P SP NT CD72 P P P SP NT TPST1 P P SP SP P GPR21 P P P SP NT USP21 SP SP P P NT FZD4 N N P SP NT TM7SF1 P P P SP NT FXYD5 N N SP SP NT RIT1 P P P SP SP CASP10 SP SP P N NT P: positive response in the assay SP: Strong positive response in the assay NT: not tested N: negative response in the assay
Expression Analysis of the TARGETS Identified in Human Primary Synovial Fibroblasts Derived from Synovium of RA Patients
 Expression levels for all the TARGETS identified are determined in at least three different isolates of primary human synovial fibroblasts.
 One isolate of RASF's is obtained as cryopreserved passage 2 cells from Cell Applications Inc. (Cat. No. 404-05). These cells are cultured and propagated in DMEM (Invitrogen) supplemented with 10% (v/v) heat-inactivated FBS (ICN) and 1×Pen/Strep (Invitrogen).
 Two other isolates are established starting from synovial membrane biopsy specimens obtained during knee arthroscopy of patients who are diagnosed as suffering from RA. Upon removal, the tissue samples are frozen in DMEM (Invitrogen) containing 15% (v/v) heat-inactivated FBS, 1× sodium pyruvate (Invitrogen), 1× antibiotics (Invitrogen) and 10% (v/v) DMSO (Sigma) and stored in liquid nitrogen. Cell culture is initiated from these synovial tissue specimens as follows: the tissues are washed thoroughly with Hanks balanced salt solution (Invitrogen) supplemented with 2× antibiotics and are digested overnight at 37° C. with 0.2% (w/v) Type IV Collagenase (Invitrogen) in DMEM containing 10% (v/v) heat-inactivated FBS, 1× sodium pyruvate, 2× antibiotics. Cells are collected, washed, resuspended in growth medium (DMEM supplemented with 10% heat-inactivated FBS, 1× sodium pyruvate, 1× antibiotics) and finally plated in 3 different wells of a 6-wells tissue culture plate. Non-adherent cells are removed after 3 days by changing growth medium. When cells reached 90-95% confluency, they are harvested by trypsinization (0.25% trypsin/1 mM EDTA) and passaged to a 25-cm2 tissue culture flask. Further passaging is done by 1/3 splitting and growth medium is changed twice a week. For expression analysis, cells are used at passages 6 to 10.
 For RNA preparation, the primary human synovial fibroblasts are seeded in 10-cm Petri dishes (500,000 cells/dish). After overnight incubation, medium is refreshed to 6 ml of M199 medium supplemented with 1% (v/v) heat-inactivated FBS containing 1× Pen/Strep. 24 h later, total RNA is extracted using the `SV Total RNA Isolation kit` (Promega). Certain samples are stimulated before harvesting. In this case, the following medium is added to the dishes for 24 h before harvesting: supernatant of THP1 cells (a human monocytic cell line) triggered with recombinant human TNFα (25 ng/ml) for 72 h in M199 medium+1% FBS diluted 2 fold in fresh M199+1% FBS.
 The concentration of RNA in each sample is fluorimetrically quantified using the `Ribogreen RNA quantitation kit` (Molecular Probes). A similar amount of RNA from each preparation is reverse transcribed into first strand cDNA with the `Taqman reverse transcription kit` from Applied Biosystems. Briefly, 40 ng RNA is included per 20 μl reaction mix containing 50 pmol of random hexamers, 10 U Rnase inhibitor, 25 U Multiscribe reverse transcriptase, 5 mM MgCl2 and 0.5 mM of each dNTP. The reaction mixture is incubated at 25° C. for 10 min, followed by 30 min incubation at 48° C. and heat inactivation (5 min 95° C.) of the reverse transcriptase in a thermocycler (Dyad, MJ Research). Reactions are immediately chilled to 4° C. at the end of the program. To avoid multiple freeze/thaw cycles of the obtained cDNA, the different samples are pooled in 96-well plates, aliquoted and stored at -20° C.
 Real-time PCR reactions are performed and monitored using the `ABI PRISM 7000 Sequence Detection System Instrument` (Applied Biosystems). Primers are designed with `Primer Express software version 2.0` (Applied Biosystems) and purchased from Sigma-Genosys. The specificity of the primers is confirmed by BLASTN searches. The PCR mixture consisted of 1× Sybr Green PCR Master mix (Aplied Biosystems), 7.5 pmol of forward and reverse primers and 2 μl of the retrotranscription reaction product in a total volume of 25 μl. After an initial denaturation step at 95° C. for 10 min, the cDNA products are amplified with 40 cycles consisting of 95° C. for 15 s and 60° C. for 1 min, followed by a dissociation protocol, which is defined as a slow ramp from 60 to 95° C. Using the dissociation protocol single peaks are confirmed in each of the PCR reactions for the various genes to exclude non-specific amplification. In order to normalize for variability in the initial quantities of cDNA between different samples, amplification reactions with the same cDNA are performed for the housekeeping genes β-actin/18S rRNA using either home made β-actin primers and SYBR Green PCR Master Mix or the `predeveloped primer and Taqman probe mix` for human 18S rRNA and `Taqman Universal PCR Mastermix no AmpErase UNG` (all Applied Biosystems) according to the manufacturer's instruction. To identify any contamination resulting from residual genomic DNA, real-time PCR reactions with product from a control (-RT) reverse transcription reaction that is performed under the same conditions but without the addition of the reverse transcriptase are included for each sample. Threshold cycle values (Ct), i.e. the cycle number at which the amount of amplified gene of interest reached a fixed threshold are determined for each sample. For each sample, the Ct value is determined by subtracting the Ct value of the endogenous control (β-actin) from the Ct value obtained for the target gene. A gene is considered as expressed in primary human SFs if the Ct value obtained for this hit is lower as 13.3 in at least one of the 3 synovial isolates, activated or not, that are available. The results of the expression profiling experiments are summarized in Table 5. The DCt value relative to β-actin obtained for all target genes (listed in Table 1) in untriggered SFs or SFs triggered with 25% `complex cytokine mixture` are given in this Table 5. The primers used in this study are listed in Table 2.
TABLE-US-00013 TABLE 5 Expression of target genes in primary synovial fibroblasts Untriggered Triggered Gene symbol RASFs RASFs RIPK2 6.7 3.7 PRKCE 8.8 7.8 MST3 6.4 5.2 MAPKAPK5 7.5 6.0 MKNK1 5.9 5.6 CAMK4 14.2 11.6 SEPT1 7.0 7.1 PGPEP1 8.7 8.1 CD72 9.0 9.1 TPST1 5.1 3.1 GPR21 11.5 9.8 USP21 8.1 6.9 FZD4 7.4 7.3 TM7SF1 7.6 7.1 FXYD5 2.8 2.1 RIT1 6.5 4.4 CASP10 14.5 11.9
Testing of the TARGETS Identified Using siRNA Technology
 When the adenoviral expression or the activation of a factor in SFs increases the collagen degrading potency of these cells, activation of this factor is sufficient to increase collagen degradation by these cells. This indicates that the factor controls or is acting on signaling pathways that are important for the regulation of MMP1 and/or other proteases involved in collagen degradation. However, to confirm that a factor is indispensable for the expression of MMP1 or degradation of collagen, the following "reverse MMP1 assay" experiments are performed. These experiments are key in determining whether the inhibition of a TARGET protein will reduce the cytokine-induced MMP1 expression, collagen degradation and thus has therapeutic potential for diseases involving ECM degradation.
 This assay used multiple "knock down" viruses corresponding to the TARGET genes that, when overexpressed or activated in SFs, increase the potency of these cells to express MMP1 or to degrade collagen. Certain "knock down" viruses are also designed against 3 other target genes (MAPKAPK5, FXYD5 and FZD4) that are not identified through the screening of the FlexSelect collection in the "MMP assay". A "knock down" virus is defined as an adenovirus that drives the expression of a self-complementing single-stranded siRNA molecule polynucleotide, resulting in the reduction of the corresponding mRNAs levels that encode the target polypeptides. The siRNA polynucleotides are designed based on the sequence of the gene encoding the TARGET polypeptide and selected according to siRNA designing rules that give an improved reduction of the target sequence expression compared to nucleotide sequences that do not comply with these siRNA designing rules (See PCT/EP03/04362). Multiple viruses are generated and tested for each TARGET gene as not every siRNA is as efficient in reducing the mRNA levels for a given TARGET gene.
 SFs are seeded in 384 or 96 well plates and infected at various MOI's with the knockdown viruses generated against the targets identified as players modulating SF MMP1 expression in, or SF collagen degradation. Five days after infection, at the time the levels of the target mRNA in the SFs are efficiently reduced by the knock down virus, the SFs are "activated" with a trigger or a mixture of triggers relevant in the field of arthritis. In uninfected SFs, or SFs infected with control knock down viruses, this trigger or mix of triggers lead to an increase in the expression of MMP1 and the potency of the cells to degrade collagen.
 Two days after application of the trigger, the levels of MMP1 in the supernatant of the SFs are measured in an MMP1 ELISA, or the degradation of collagen by the supernatant of the SFs is measured in the collagen degradation assay. If the reduction in the expression level for a certain target gene leads to a reduced response of the cells to the RA-relevant trigger applied, this indicates that this target gene is indispensable for the SFs to respond to this trigger. The inhibition of the activity of the polypeptide product of this gene, or the reduction in expression of this gene, might thus represent a suitable approach for treatment of RA.
 In order to work in an unbiased way, a complex mixture of factors relevant in the field of RA is generated as follows: THP-1 cells, a representative human monocyte cell line, is cultured in the presence of human recombinant TNFalpha (Sigma, 25 ng/ml) for 48 h. Supernatant of this cell line is then collected and stored at -80° C. until further use. The monocytes respond to the TNF-alpha trigger by the production of a variety of other cytokines and factors, most of which will be pro-inflammatory. As monocytes (macrophages) as well as high levels of TNF-alpha are present in the affected joints of RA patients, the trigger mixture produced in this way is relevant in the field of RA and will be representative for the mixture of factors present in the joints of RA patients. The unbiased character of this method represents an important advantage, as the mixture produced is very complex and might contain factors unknown to be involved in RA or even factors unknown to date.
 The white bars in FIG. 10 show the increase of SF MMP1 expression upon treatment with cytokines relevant in the field of RA (TNF, IL1 and OSM) or a combination thereof. For this experiment, SFs are seeded in 96 well plates, 3,000 cells/well. 24 h later, the medium is changed to M199 medium supplemented with 1% FBS. One day after the medium change, cytokines or combinations thereof are added to the cultures, each cytokine being added to a final concentration of 25 ng/ml. 72 h after cytokine addition, the supernatant is collected and processed in the MMP1 ELISA. In parallel with this experiment, SFs are triggered, using the same protocol, with the supernatant of THP1 cells (2-fold diluted in M199+1% FBS) that are left untreated or are treated with the same cytokines or combinations of cytokines for 48 h in M199 medium+1% FBS. MMP1 levels for these samples are shown in FIG. 10 as grey bars. The induction of the MMP1 expression levels by the supernatants of TNF-treated THP1 cells is stronger (>4.5 fold induction) as compared to the induction by recombinant TNF alone (3-fold induction) and almost equals the 5-fold induction obtained by a mixture of 3 purified cytokines (TNFalpha, IL1b, OSM). This result indicates that the supernatant of TNF-induced THP1 cells contains additional pro-inflammatory factors that trigger the SFs towards MMP1 production.
 In another experiment, inhibition of the response of SFs to the SN (supernatant) of TNF-triggered THP1 cells is investigated. SFs are seeded in 384 well plates at 1500 cells/well and left uninfected or infected with the control knock-down virus Ad5-eGFP KD or the control knock-in virus Ad5-MMP1. One day after infection, dexamethasone, a classical anti-inflammatory agent and SB203580 (an inhibitor of p38alpha and p38beta (kinases involved in the response of cells to TNF and other cytokines), purchased at Calbiochem, dissolved in 100% DMSO), are added to the SF cultures at a final concentration of 100 nM and 5 μM respectively, 1 h before triggering of the cells with 2-fold diluted SN of TNF-activated THP1 cells. 72 h after treatment, the SN is collected and subjected to the MMP1 ELISA. Results are depicted in FIG. 11: SFs are left uninfected or are infected with a control knock-in virus (Ad5-MMP1_KI) or a control knock-down virus (Ad5-eGFP KD). Raw luminescence signals, which are proportional to the MMP1 levels, are shown.
 Triggering of the cells led to a 6-fold increase of MMP1 expression. Even higher MMP1 levels are measured in the samples infected with Ad5-MMP1, indicating that the THP1 SN-induced MMP1 levels are not saturating for the MMP1 ELISA. The MMP1 levels obtained in the dexamethasone and SB203580 treated samples are 4 and 3 fold lower as the control levels, respectively, indicating that the assay as set up is suitable for the identification of inhibitors of the inflammatory response of SFs. Efficient reduction of gene expression in SFs can be obtained by RNAi (RNA interference) using knockdown viruses or transfection of siRNA duplexes.
Analysis of the Reduction in mRNA Expression of TARGET Genes by Ad-siRNA
 Primary human synovial fibroblasts are seeded in gelatin coated 6-well plates (75,000 cells/well) in 2 ml synovial growth medium (Cell Applications Inc.) supplemented with 1×Pen/Strep (Invitrogen). After overnight incubation, cells are infected with the Ad5-siRNA targeting the gene of interest at an MOI of 3000. As a negative control, other wells are infected at the same MOI with Ad5-siRNA targeting the firefly luciferase gene. Five days post infection, medium is refreshed with 2 ml M199 medium supplemented with 1% (v/v) heat-inactivated FBS. At the same time, parallel samples are stimulated by refreshing the medium with 2 ml of a 2-fold dilution of the `complex cytokine mixture` in M199+1% FBS. 48 h later, total RNA is extracted using the ` SV Total RNA Isolation kit` (Promega). RNA is quantitated and cDNA is prepared as described in Example 5. For each sample, real-time PCR reactions are performed for the TARGET and the 18S rRNA genes and Ct values are calculated as previously described in Example 5. To calculate the % knock-down of the endogenous TARGET mRNA after infection with the Ad5-siRNA, values are first expressed relative to the control samples that are infected with Ad5-luciferase-v13_KD virus using the equation: relative expression=2Ct with Ct=Ct.sub.(sample infected with Ad5-luciferase-v13--KD)-Ct.sub.(sample infected with TARGETspecific Ad5-siRNA). The DCt values indicate the expression relative to β-actin as indicated in Example 5. Table 6 shows that after infection with most of the selected Ad5-siRNAs, more than 60% reduction of the TARGET mRNA, irrespective of whether the cells are stimulated with the `complex cytokine mixture`. The abbreviation "Rel Expr" means relative expression.
TABLE-US-00014 TABLE 6 no trigger triggered TARGET Ad5-siRNA DCt DDCt Rel. Expr. % KD DCt DDCt Rel. Expr. % KD CAMKIV Ad5-CamK4-v1_KD 16.7 -2.9 0.13 86.6 18.3 -3.8 0.07 92.8 Ad5-Luciferase-v13_KD 13.8 0 1.00 0.0 14.5 0 1.00 0.0 PRKCE Ad5-PRKCE-v11_KD 10 -1.1 0.47 53.3 8.7 -0.9 0.54 46.4 Ad5-Luciferase-v13_KD 8.9 0 1.00 0.0 7.8 0 1.00 0.0 MMP1 Ad5-MMP1-v10_KD 13.4 -4.9 0.03 96.7 5 -3.2 0.11 89.1 Ad5-Luciferase-v13_KD 8.5 0 1.00 0.0 1.8 0 1.00 0.0 MAPKAPK5 Ad5-MAPKAPK5-v2_KD 9.3 -2.3 0.20 79.7 7.4 -3 0.13 87.5 Ad5-MAPKAPK5-v8_KD 9.2 -2.2 0.22 78.2 7.3 -2.9 0.13 86.6 Ad5-Luciferase-v13_KD 7 0 1.00 0.0 4.4 0 1.00 0.0 RIT Ad5-RIT-v5_KD 7.1 -1.2 0.44 56.5 6.5 -1.9 0.27 73.2 Ad5-Luciferase-v13_KD 5.9 0 1.00 0.0 4.6 0 1.00 0.0 TPST1 Ad5-TPST1-v1_KD 7.3 -0.9 0.54 46.4 8.2 -2.3 0.20 79.7 Ad5-Luciferase-v13_KD 6.4 0 1.00 0.0 5.9 0 1.00 0.0 USP21 Ad5-USP21-v3_KD 9.5 -1.2 0.44 56.5 8.9 -1.3 0.41 59.4 Ad5-Luciferase-v13_KD 8.3 0 1.00 0.0 7.6 0 1.00 0.0 MST3 Ad5-MST3-v4_KD 6.9 -2 0.25 75.0 7.1 -2.1 0.23 76.7 Ad5-STK24-v1_KD 7.8 -2.9 0.13 86.6 6.4 -1.4 0.38 62.1 Ad5-Luciferase-v13_KD 4.9 0 1.00 0.0 5 0 1.00 0.0
Ad-siRNA Viruses Function to Knock Down Expression of MAPKAPK5, PRKCE and CAMK4 at the Protein Level
 FIG. 14 illustrates the functionality of Ad-siRNAs for reducting expression of TARGET genes (PRKCE, MAPKAPK5 and CAMK4) at the protein level in human cells.
 Recombinant adenoviruses mediating the expression of siRNA's targeting MAPKAPK5, PRKCE and CAMK4 are generated according to the procedure described in WO03/020931. The target sequences in these genes based on which the siRNAs were designed and that were used to generate the recombinant adenoviruses are listed in Table 3.
 The functionality of MAPKAPK5 targeting adenoviruses is tested as follows: On day 1,500.000 primary human SFs are seeded per petri dish. One day later, the cells are infected with Ad5-MAPKAPK5-v2_KD, Ad5-MAPKAPK5-v8_KD or Ad5-eGFP-v5_KD at an MOI of 4000 (based on the titers (number of virus particles per ml) defined for the viruses by Q-rt-PCR). On day 7, cells are detached from the petri dish according to standard procedure using a trypsin EDTA solution. The trypsin is then neutralized by addition of DMEM growth medium supplemented with 10% FBS. The cells are then collected by a centrifugation step (1000 rpm, 5 min). The pellet is lysed in 100 μl of fresh RIPA buffer (50 mM Tris pH7.5, 150 mM NaCl, 1% deoxycholate, 1% Triton X100, 0.1% SDS). The samples are then sonicated for 10 sec. The protein concentration of the samples is then determined using the BCA kit (Pierce, Cat No 23227) as described by the provider, using BSA as a standard. To 30 μg of cell lysate diluted to 19.5 μl in RIPA buffer, 3.5 μl of reducing agent (NuPage reducing agent No 10, Invitrogen NP0004) and 7.5 μl of sample buffer (NuPage LDS sample buffer, Invitrogen NP0007) are added. The 30 μl sample is then boiled for 5 min and loaded on a 10% polyacrylamide gel (Invitrogen NP0301). The gel is then run for 2 hours at 100V in 1×MOPS/SDS NuPage running buffer (Invitrogen NP001). 10 μl of Seablue Plus Prestained standard (Invitrogen LC5925) is used to estimate protein size on the gel. The proteins on the gel are then transferred onto a PVDF membrane (Invitrogen LC2002) by a wet blotting procedure using a transfer buffer prepared by mixing 100 ml Nupage Transfer buffer 20* (NP0006-1), 400 ml methanol and 1500 ml Milli Q water. Before the transfer, the membrane is first soaked in methanol and in transfer buffer. The transfer is performed at 100V for 90 minutes. The membrane is then blocked by 30 min soaking in blocking buffer (2% blocking powder (Amersham, RPN 2109) prepared in PBST (PBS supplemented with 0.1% Tween 20 (Sigma, P1379)). After blocking, the immunodetection is performed using a mouse monoclonal antibody against MAPKAPK5 (BD Biosciences, Cat No 612080) diluted 250 fold in blocking buffer. After overnight incubation with this primary antibody, the membrane is washed 3 times with PBST and incubated 1 hr with the secondary antibody ((Polyclonal goat anti-mouse Ig, HRP conjugated (DAKO P0447) diluted 50000 fold in blocking buffer. The blot is then washed 3 times in PBST and the detection is performed with ECL advance (RPN2109, Amersham) on a Kodakimager according to the manufacturers instructions. The Western Blotting revealed a lower expression level of MAPKAPK5 in the Ad5-MAPKAPK5-v2_KD and Ad5-MAPKAPK5-v8_KD infected cells compared to the cells infected with the Ad5-eGFP-v5_KD negative control virus. Equal loading of the 30 μg samples is demonstrated by immunodetection of β-actin after removal of the MAPKAPK5 antibody by a `stripping procedure` (5 minutes boiling of the membrane in PBST). Immunodetection of β-actin is performed according to the method described for MAPKAPK5 detection, but using a goat polyclonal antibody against β-actin (Santa Cruz, Cat No SC-1615) at a 1000 fold dilution as primary antibody and a rabbit anti goat antibody at a 50000 fold dilution as a secondary antibody. Results of this experiment are shown in FIG. 14 C.
 The functionality of the PRKCE targeting adenovirus (Ad5-PRKCE-v11_KD) is tested according to the same protocol as the one described above for MAPKAPK5, with the difference that an MOI of 2000 is used for infection of the cells. The western blotting procedure is the same as the one described for MAPKAPK5 detection, with the difference that a PRKCE specific antibody is used (BD Biosciences, Cat No 610085) at a dilution of 250-fold. The same secondary antibody is used as for the detection of MAPKAPK5. Results are shown in FIG. 14 B.
 The functionality of the CAMK4 targeting adenovirus is tested as follows: These adenoviruses are used to infect Hek293T cells cultured in 6-well plates as follows. On day 1, 400000 Hek293T cells are seeded per 6-well plate in DMEM+10% FBS. One day later, the cells are infected with Ad5-CAMK4-v1_KD, CAMK4-CAMK4-v9_KD or Ad5-eGFP-v5_KD at an MOI (multiplicity of infection) of 500 (based on the titers (number of virus particles per ml) defined for the viruses by Q-rt-PCR). One day after the infection, the medium is refreshed. On day 7, cells are detached from the petri dish according to standard procedure using a trypsin EDTA solution. The handling of the cell pellet, the running/blotting of the gel and the immunodetection procedure is identical to what is described for MAPKAPK5, with the difference that 40 μg protein is loaded on the gel and that a mouse monoclonal antibody against CAMK4 (Santa Cruz, Sc-17762, diluted 100-fold in blocking buffer) is used. The Western Blotting reveals a lower expression level of CAMK4 in the Ad5-CAMK4-v1_KD and the Ad5-CAMK4-v9_KD infected cells compared to the cells infected with the Ad5-eGFP-v5_KD negative control virus. Equal loading of the 30 μg samples is demonstrated by immunodetection of β-actin after removal of the CAMK4 antibody by a `stripping procedure`. Results of this experiment are given in FIG. 14 A.
 These experiments demonstrate that the Ad-siRNA virus function to reduce the expression levels of the corresponding MAPKAPK5, CAMK4 and PRKCE polpeptides in human cells.
Reduction of the Expression in Primary SFs of Various TARGET Genes by Ad-siRNAs Inhibit SF-Induced MMP1 Expression
 FIG. 12 illustrates the reduction of cytokine-induced SF MMP1 expression by Ad-siRNAs reducing the expression of TARGET genes. These Ad-siRNAs are generated according to the procedure described in WO03/020931. The target sequences (KD SEQ) in these genes, based on which the siRNAs were designed and that were used to generate the recombinant adenoviruses, are listed in Table 3.
 The efficacy of Ad5-siRNAs in the `MMP assay` is tested as follows. Day 1, SFs (passage 9 to 10) are seeded in 96 well plates at a density of 3000 cells per well in complete synovial growth medium (Cell Applications). One day later, the cells are infected with increasing amounts (3, 7.5, 12 or 15 μl in experiment shown in FIG. 12 A; 3, 6, 9, 12 and 15 μl in experiment shown in FIG. 12 B; and 3, 6, 9, and 12 μl in the experiments represented on FIGS. 12 C and 12 D) of the Ad-siRNA's. The following viruses are used as negative control: Ad5-eGFP-v5_KD, Ad5-Luciferase-v13_KD and Ad-M6PR-v1_KD. Ad5-MMP1-v10_KD is used as a positive control virus. The virus load is corrected by addition of the neutral virus Ad5-Luciferase-v13_KD to bring the final virus volume on the cells to 15 μl in every well. This correction guarantees that the effects observed do not result from differences in the virus load applied to the cells. The cells are then incubated for 5 days before the activation step. This step involves the replacement, in every well, of the growth medium by 75 μl of M199 medium supplemented with 25 μl of `complex trigger`. 48 hrs after the activation step, the supernatant is collected and subjected to the MMP1 ELISA as described above.
 The results of the experiment are shown in FIG. 12A, B, C and D. The average of duplicate data points is shown in these Figures. The quality of the experiment is demonstrated by the efficacy of the Ad-siRNA virus targeting MMP1 itself. This positive control virus strongly reduces the MMP1 expression by SFs, whereas the negative control viruses, designed to target the expression of luciferase, M6PR and eGFP do not influence the levels of MMP1 expression, as expected. The Ad-siRNAs designed against TARGET genes (GPR21, FZD4, TM7SF1, PGPEP1, SEPT1, CD72, FXYD5 (FIG. 12 A.); PRKCE, CAMK4, MAPKAPK5 (FIG. 12 B.), RIPK2, RITZ (FIG. 12 C.) and PPST1, USP21 and STK24 (FIG. 12 D.), also lead to a clear reduction of the complex trigger induced MMP1 expression by primary human SFs. For certain TARGET genes (e.g. CAMK4, MAPKAPK5), 2 independent Ad-siRNAs showed efficacy in reducing cytokine induced MMP1 expression by SFs. In FIGS. 12 A and B, the MMP1 expression levels are shown in terms of raw data (RLU) whereas in FIGS. 12 C and 12 D, the MMP1 expression levels are expressed relative to the samples infected with Ad5-luciferase-v13_KD only set to 100%.
 For most TARGET genes, at least 1 of the 5 Ad-siRNAs designed per TARGET gene mediated a reduction of the cytokine-induced MMP1 expression by SFs. This was not the case for MKNK1 and CASP10. The effects observed were weaker for USP21 and MST3.
 It can be concluded, from this experiment, that these genes represent valuable drug targets that are shown to modulate MMP1 expression in SFs. Similarly, the inhibition of the activity of the protein product of these genes by a small molecule compound is expected to reduce the `complex cytokine` induced MMP1 expression in the `MMP assay`. The inhibition of the activity of the protein products of these genes by small molecule compounds is also predicted to reduce the degradation of the joint associated with RA.
Reduction of the Expression in Primary SFs of MAPKAPK5 and CAMK4 by Ad-siRNAs Inhibit Cytokine-Induced Collagen Degradation
 This experiment measures the ability of Ad-siRNAs to reduce cytokine-induced degradation of collagen type I, which is even more stringent than the MMP1 ELISA, as the degradation of native collagen might be due to the action of proteases different from MMP1. The Ad-siRNAs used in this experiment are generated according to the procedure described in WO03/020931. The recombinant Ad-siRNAs used in this experiment were generated based on target sequences in the target genes that are listed in Table 3.
 The efficacy of Ad5-siRNAs in the `miniaturized native collagen type I degradation assay` described above is tested as follows: Day 1, SFs (passage 9 to 10) are seeded in 96 well plates at a density of 3000 cells per well in complete synovial growth medium (Cell Applications). One day later, the cells are infected with increasing amounts (3, 6, 9, 12 and 15 μl) of the Ad-siRNA's indicated on the figure. The following viruses are used as negative control: Ad5-eGFP-v5_KD, and Ad5-Luciferase-v13_KD. The virus load is corrected by addition of the neutral virus Ad5-Luciferase-v13_KD to bring the final virus volume added to each well to 15 μl. This correction guarantees that the effects observed do not result from differences in the virus load applied to the cells. The cells are then incubated for 5 days before the activation step. This step involves the replacement, in every well, of the growth medium by 45 μl of M199 medium supplemented with 15 μl of `complex trigger`. 4 days later, the supernatant is collected and subjected to the miniaturized collagen type I degradation assay according to the protocol as described above. The results of the experiment are shown in FIG. 15.
 The negative control viruses, designed to target the expression of luciferase and eGFP, do not influence the levels of collagen degradation, as expected. The Ad-siRNAs targeting MAPKAPK5 and CAMK4 do mediate a clear reduction of the complex trigger-induced collagen degradation by primary human SFs. It can be concluded, from this experiment, that these genes represent valuable drug targets that are shown to modulate collagen degradation by SFs. Similarly, the inhibition of the activity of the protein product of these genes by a small molecule compound is expected to reduce the `complex cytokine` induced collagen degradation by SFs. The inhibition of the activity of the protein products of these genes by small molecule compounds is also predicted to reduce the degradation of the joint associated with RA. In similar experiments, the Ad5-MMP1-v10_KD virus is shown to strongly reduce the cytokine induced collagen degradation by SFs, which implies the fact that MMP1 itself is the main collagenase responsible for the cytokine induced collagen degradation by SFs. As such, this means that modulation of MMP1 expression by SFs is sufficient to reduce cartilage degradation associated with RA.
Identification of Small Molecules that Inhibit Target Kinase Activity
 Compounds are screened for inhibition of the activity of the TARGETS that are kinase polypeptides. The affinity of the compounds to the polypeptides is determined in an experiment detecting changed reaction conditions after phosphorylation. The TARGET kinase polypeptides are incubated with its substrate and ATP in an appropriate buffer. The combination of these components results in the in vitro phosphorylation of the substrate. Sources of compounds include commercially available screening library, peptides in a phage display library or an antibody fragment library, and compounds that have been demonstrated to have binding affinity for a TARGET kinase.
 The TARGET kinase polypeptides can be prepared in a number of ways depending on whether the assay will be run using cells, cell fractions or biochemically, on purified proteins. The polypeptides can be applied as complete polypeptides or as polypeptide fragments, which still comprise TARGET kinase catalytic activity.
 Identification of small molecules inhibiting the activity of the TARGET kinase polypeptides is performed by measuring changes in levels of phosphorylated substrate or ATP. Since ATP is consumed during the phosphorylation of the substrate, its levels correlate with the kinase activity. Measuring ATP levels via chemiluminescent reactions therefore represents a method to measure kinase activity in vitro (Perkin Elmer). In a second type of assay, changes in the levels of phosphorylated substrate are detected with phosphospecific agents and are correlated to kinase activity. These levels are detected in solution or after immobilization of the substrate on a microtiter plate or other carrier. In solution, the phosphorylated substrate is detected via fluorescence resonance energy transfer (FRET) between the Eu labeled substrate and an APC labeled phosphospecific antibody (Perkin Elmer), via fluorescence polarization (FP) after binding of a phosphospecific antibody to the fluorescently labeled phosphorylated substrate (Panvera), via an Amplified Luminescent Proximity Homogeneous Assay (ALPHA) using the phosphorylated substrate and phosphospecific antibody, both coupled to ALPHA beads (Perkin Elmer) or using the IMAP binding reagent that specifically detects phosphate groups and thus alleviates the use of the phosphospecific antibody (Molecular Devices). Alternatively, the substrate is immobilized directly or by using biotin-streptavidin on a microtiter plate. After immobilization, the level of phosphorylated substrate is detected using a classical ELISA where binding of the phosphospecific antibody is either monitored via an enzyme such as horseradish peroxidase (HRP) or alkaline phosphatase (AP) which are either directly coupled to the phosphospecific antibody or are coupled to a secondary antibody. Enzymatic activity correlates to phosphorylated substrate levels. Alternatively, binding of the Eu-labeled phosphospecific antibody to the immobilized phosphorylated substrate is determined via time resolved fluorescence energy (TRF) (Perkin Elmer). In addition, the substrate can be coated on FLASH plates (Perkin Elmer) and phosphorylation of the substrate is detected using 33P labeled ATP or 125I labeled phosphospecific antibody.
 Small molecules are randomly screened or are preselected based upon drug class, (i.e. known kinase inhibitors), or upon virtual ligand screening (VLS) results. VLS uses virtual docking technology to test large numbers of small molecules in silico for their binding to the polypeptide of the invention. Small molecules are added to the kinase reaction and their effect on levels of phosphorylated substrate is measured with one or more of the above-described technologies.
 Small molecules that inhibit the kinase activity are identified and are subsequently tested at different concentrations. IC50 values are calculated from these dose response curves. Strong binders have an IC50 in the nanomolar and even picomolar range. Compounds that have an IC50 of at least 10 micromol or better (nmol to pmol) are applied in alkaline phosphatase assay or bone mineralization assay to check for their effect on the induction of osteogenesis.
Ligand Screens for Target GPCRs
Reporter Gene Screen.
 Mammalian cells such as Hek293 or CHO-K1 cells are either stably transfected with a plasmid harboring the luciferase gene under the control of a cAMP dependent promoter (CRE elements) or transduced with an adenovirus harboring a luciferase gene under the control of a cAMP dependent promoter. In addition reporter constructs can be used with the luciferase gene under the control of a Ca2+ dependent promoter (NF-AT elements) or a promoter that is controlled by activated NF-κB. These cells, expressing the reporter construct, are then transduced with an adenovirus harboring the cDNA of a TARGET GPCR. Forty (40) hours after transduction the cells are treated with the following:
 a) an agonist for the receptor and screened against a large collection of reference compounds comprising peptides (LOPAP, Sigma Aldrich), lipids (Biomol, TimTech), carbohydrates (Specs), natural compounds (Specs, TimTech), small chemical compounds (Tocris), commercially available screening libraries, and compounds that have been demonstrated to have binding affinity for a polypeptide comprising an amino acid sequence selected from the group consisting of the SEQ ID NOs of the TARGET GPCRs; or
 b) a large collection of reference compounds comprising peptides (LOPAP, Sigma Aldrich), lipids (Biomol, TimTech), carbohydrates (Specs), natural compounds (Specs, TimTech), small chemical compounds (Tocris), commercially available screening libraries, and compounds that have been demonstrated to have binding affinity for a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs of the TARGET GPCRs.
 Compounds, which decrease the agonist induced increase in luciferase activity or the constitutive activity, are considered to be antagonists or inverse agonists for a TARGET GPCR. These compounds are screened again for verification and screened against their effect on osteoblast differentiation. The compounds are also screened to verify binding to the GPCR. The binding, osteogenesis and reporter activity assays can be performed in essentially any order to screen compounds.
 In addition, cells expressing the NF-AT reporter gene can be transduced with an adenovirus harboring the cDNA encoding the α-subunit of G15 or chimerical Gα subunits. G15 is a promiscuous G protein of the Gq class that couples to many different GPCRs and as such re-directs their signaling towards the release of intracellular Ca2+ stores. The chimerical G alpha subunits are members of the Gs and Gi/o family by which the last 5 C-terminal residues are replaced by those of G.sub.αq, these chimerical G-proteins also redirect cAMP signaling to Ca2+ signaling.
 Mammalian cells such as Hek293 or CHO-K1 cells are stably transfected with an expression plasmid construct harboring the cDNA of a TARGET GPCR. Cells are seeded, grown, and selected until sufficient stable cells can be obtained. Cells are loaded with a Ca2+ dependent fluorophore such as Fura3 or Fura4. After washing away the excess of fluorophore the cells are screened against a large collection of reference compounds comprising peptides (LOPAP, Sigma Aldrich), lipids (Biomol, TimTech), carbohydrates (Specs), natural compounds (Specs, TimTech), small chemical compounds (Tocris), commercially available screening libraries, and compounds that have been demonstrated to have binding affinity for a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs of the TARGET GPCRs, by simultaneously adding an agonist (alternatively no agonist need be added if the constitutive activity of the receptor is used) and a compound to the cells. Activation of the receptor is measured as an almost instantaneously increase in fluorescence due to the interaction of the fluorophore and the Ca2+ that is released. Compounds that reduce or inhibit the agonist induced increase in fluorescence (or constitutive fluorescence) are considered to be antagonists or inverse agonists for the receptor they are screened against. These compounds are screened again to measure the amount of osteoblast differentiation as well as binding to a TARGET GPCR.
 CHO cells, stably expressing Apoaequorin are stably transfected with a plasmid construct harboring the cDNA of a TARGET GPCR. Cells are seeded, grown, and selected until sufficient stable cells can be obtained. The cells are loaded with coelenterazine, a cofactor for apoaequorin. Upon receptor activation intracellular Ca2+ stores are emptied and the aequorin will react with the coelenterazine in a light emitting process. The emitted light is a measure for receptor activation. The CHO, stable expressing both the apoaequorin and the receptor are screened against a large collection of reference compounds comprising peptides (LOPAP, Sigma Aldrich), lipids (Biomol, TimTech), carbohydrates (Specs), natural compounds (Specs, TimTech), small chemical compounds (Tocris), commercially available screening libraries, and compounds that have been demonstrated to have binding affinity for a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs of the TARGET GPCRs, by simultaneously adding an agonist (alternatively no agonist need be added if the constitutive activity of the receptor is used) and a compound to the cells. Activation of the receptor is measured as an almost instantaneously light flash due to the interaction of the apoaequorin, coelenterazine, and the Ca2+ that is released. Compounds that reduce or inhibit the agonist induced increase in light or the constitutive activity are considered to be antagonists or inverse agonists for the receptor they are screened against. These compounds are screened again to measure the amount of osteoblast differentiation as well as binding to a TARGET GPCR.
 In addition, CHO cells stable expressing the apoaequorin gene are stably transfected with a plasmid construct harboring the cDNA encoding the α-subunit of G15 or chimerical G.sub.α, subunits. G15 is a promiscuous G protein of the Gq class that couples to many different GPCRs and as such redirects their signaling towards the release of intracellular Ca2+ stores. The chimerical G alpha subunits are members of the Gs and Gi/o family by which the last 5 C-terminal residues are replaced by those of G.sub.αq, these chimerical G-proteins also redirect cAMP signaling to Ca2+ signaling.
Screening for Compounds that Bind to the GPCR Polypeptides (Displacement Experiment)
 Compounds are screened for binding to the TARGET GPCR polypeptides. The affinity of the compounds to the polypeptides is determined in a displacement experiment. In brief, the GPCR polypeptides are incubated with a labeled (radiolabeled, fluorescent labeled) ligand that is known to bind to the polypeptide and with an unlabeled compound. The displacement of the labeled ligand from the polypeptide is determined by measuring the amount of labeled ligand that is still associated with the polypeptide. The amount associated with the polypeptide is plotted against the concentration of the compound to calculate IC50 values. This value reflects the binding affinity of the compound to its TARGET, i.e. the TARGET GPCR polypeptides. Strong binders have an IC50 in the nanomolar and even picomolar range. Compounds that have an IC50 of at least 10 micromol or better (nmol to pmol) are applied an osteoblast differentiation assay to check for their effect on osteogenesis. The TARGET GPCR polypeptides can be prepared in a number of ways depending on whether the assay are run on cells, cell fractions or biochemically, on purified proteins. Screening for compounds that bind to a TARGET GPCR (generic GPCR screening assay)
 When a G protein receptor becomes constitutively active, it binds to a G protein (Gq, Gs, Gi, Go) and stimulates the binding of GTP to the G protein. The G protein then acts as a GTPase and slowly hydrolyses the GTP to GDP, whereby the receptor, under normal conditions, becomes deactivated. However, constitutively activated receptors continue to exchange GDP to GTP. A non-hydrolyzable analog of GTP, [35S]GTPγS, can be used to monitor enhanced binding to membranes which express constitutively activated receptors. It is reported that [35S]GTPγS can be used to monitor G protein coupling to membranes in the absence and presence of ligand. Moreover, a preferred approach is the use of a GPCR-G protein fusion protein. The strategy to generate a TARGET GPCR-G protein fusion protein is well known for those known in the art. Membranes expressing TARGET GPCR-G protein fusion protein are prepared for use in the direct identification of candidate compounds such as inverse agonist. Homogenized membranes with TARGET GPCR-G protein fusion protein are transferred in a 96-well plate. A pin-tool is used to transfer a candidate compound in each well plus [35S]GTPγS, followed by incubation on a shaker for 60 minutes at room temperature. The assay is stopped by spinning of the plates at 4000 RPM for 15 minutes at 22° C. The plates are then aspirated and radioactivity is then read.
Receptor Ligand Binding Study On Cell Surface
 The receptor is expressed in mammalian cells (Hek293, CHO, COS7) by adenoviral transducing the cells (see U.S. Pat. No. 6,340,595). The cells are incubated with both labeled ligand (iodinated, tritiated, or fluorescent) and the unlabeled compound at various concentrations, ranging from 10 pM to 10 μM (3 hours at 4° C.: 25 mM HEPES, 140 mM NaCl, 1 mM CaCl2, 5 mM MgCl2 and 0.2% BSA, adjusted to pH 7.4). Reactions mixtures are aspirated onto PEI-treated GF/B glass filters using a cell harvester (Packard). The filters are washed twice with ice cold wash buffer (25 mM HEPES, 500 mM NaCl, 1 mM CaCl2, 5 mM MgCl2, adjusted to pH 7.4). Scintillant (MicroScint-10; 35 μl) is added to dried filters and the filters counted in a (Packard Topcount) scintillation counter. Data are analyzed and plotted using Prism software (GraphPad Software, San Diego, Calif.). Competition curves are analyzed and IC50 values calculated. If one or more data points do not fall within the sigmoidal range of the competition curve or close to the sigmoidal range the assay is repeated and concentrations of labeled ligand and unlabeled compound adapted to have more data points close to or in the sigmoidal range of the curve.
 Receptor Ligand Binding Studies on Membrane Preparations
 Membranes preparations are isolated from mammalian cells (Hek293, CHO, COS7) cells over expressing the receptor is done as follows: Medium is aspirated from the transduced cells and cells are harvested in 1×PBS by gentle scraping. Cells are pelleted (2500 rpm 5 min) and resuspended in 50 mM Tris pH 7.4 (10×106 cells/ml). The cell pellet is homogenized by sonicating 3×5 sec (UPS OH; sonotrode MS1; max amplitude: 140 μm; max Sonic Power Density: 125 W/cm2). Membrane fractions are prepared by centrifuging 20 min at maximal speed (13,000 rpm ˜15,000 to 20,000 g or rcf). The resulting pellet is resuspended in 500 μl 50 mM Tris pH 7.4 and sonicated again for 3×5 sec. The membrane fraction is isolated by centrifugation and finally resuspended in PBS. Binding competition and derivation of IC50 values are determined as described above.
Internalization Screen (1)
 Activation of a GPCR-associated signal transduction pathway commonly leads to translocation of specific signal transduction molecules from the cytoplasm to the plasma membrane or from the cytoplasm to the nucleus. Norak has developed their transfluor assay based on agonist-induced translocation of receptor-β-arrestin-GFP complex from the cytosol to the plasma membrane and subsequent internalization of this complex, which occurs during receptor desensitization. A similar assay uses GFP tagged receptor instead of β-arrestin. Hek293 cells are transduced with a TARGET GPCR vector that translates for a TARGET GPCR-eGFP fusion protein. 48 hours after transduction, the cells are set to fresh serum-free medium for 60 minutes and treated with a ligand for 15, 30, 60 or 120 minutes at 37° C. and 5% CO2. After indicated exposure times, cells are washed with PBS and fixed with 5% paraformaldehyde for 20 minutes at RT. GFP fluorescence is visualized with a Zeiss microscope with a digital camera. This method aims for the identification of compounds that inhibit a ligand-mediated (constitutive activity-mediated) translocation of the fusion protein to intracellular compartments.
Internalization Screen (2)
 Various variations on translocation assays exists using β-arrestin and β-galactosidase enzyme complementation and BRET based assays with receptor as energy donor and β-arrestin as energy acceptor. Also the use of specific receptor antibodies labeled with pH sensitive dyes are used to detect agonist induced receptor translocation to acidic lysosomes. All of the translocation assays are used for screening for both agonistic and antagonistic acting ligands.
Melanophore Assay (Arena Pharmaceutical)
 The melanophore assay is based on the ability of GPCRs to alter the distribution of melanin containing melanosomes in Xenopus melanophores. The distribution of the melanosomes depends on the exogenous receptor that is either Gi/o or Gs/q coupled. The distribution of the melanosomes (dispersed or aggregated) is easily detected by measuring light absorption. This type of assay is used for both agonist as well as antagonist compound screens.
 Andreakos E, et al. (2003). Arthritis Rheum. 48: 1901-12.  Choy E H, Panayi G S. (2001). N Engl J. Med. 344: 907-16.  Cortez-Retamozo V, et al. (2004). Cancer Res. 64(8): 2853-7  Coussens L M, et al. (2002). Science 295: 2387-92.  Creemers E E, et al. (2001). Circ Res. 2001 89:201-10  Cunnane G, et al. (2001). Arthritis Rheum 44: 2263-74.  Firestein G S. (2003). Nature. 423:356-61.  Gapski R, et al. (2004). J. Periodontol. 75:441-52.  Gomez-Reino J J, et al. (2003). Arthritis Rheum. 48: 2122-7.  Lee D M, Weinblatt M E (2001). Lancet. 358: 903-11.  Lipinski C A, et al. (2001). Advanced Drug Delivery Reviews. 46(1-3): 3-26.  Maini R N, et al. (2004). Arthritis Rheum. 50: 1051-65.  Rosenberg G A. (2002). Glia. 39:279-91.  Schanstra J P, et al. (2002). J Clin Invest. 110:371-9.  Smolen J S, Steiner G. (2003). Nat Rev Drug Discov. 2: 473-88.  Suzuki R, et al. (2004). Treat Respir Med. 3:17-27.  Vincenti M P, Brinckerhoff C E. (2002). Arthritis Res 4:157-64
 It will be appreciated by those skilled in the art that the foregoing description is exemplary and explanatory in nature, and is intended to illustrate the invention and its preferred embodiments. Through routine experimentation, an artisan will recognise apparent modifications and variations that may be made without departing from the spirit of the invention. Thus, the invention is intended to be defined not by the above description, but by the following claims and their equivalents.
29612521DNAHomo sapiens 1gaagctcttt cgcggcgcta cggcgttggc accagtctct agaaaagaag tcagctctgg 60ttcggagaag cagcggctgg cgtgggccat ccggggaatg ggcgccctcg tgacctagtg 120ttgcggggca aaaagggtct tgccggcctc gctcgtgcag gggcgtatct gggcgcctga 180gcgcggcgtg ggagccttgg gagccgccgc agcagggggc acacccggaa ccggcctgag 240cgcccgggac catgaacggg gaggccatct gcagcgccct gcccaccatt ccctaccaca 300aactcgccga cctgcgctac ctgagccgcg gcgcctctgg cactgtgtcg tccgcccgcc 360acgcagactg gcgcgtccag gtggccgtga agcacctgca catccacact ccgctgctcg 420acagtgaaag aaaggatgtc ttaagagaag ctgaaatttt acacaaagct agatttagtt 480acattcttcc aattttggga atttgcaatg agcctgaatt tttgggaata gttactgaat 540acatgccaaa tggatcatta aatgaactcc tacataggaa aactgaatat cctgatgttg 600cttggccatt gagatttcgc atcctgcatg aaattgccct tggtgtaaat tacctgcaca 660atatgactcc tcctttactt catcatgact tgaagactca gaatatctta ttggacaatg 720aatttcatgt taagattgca gattttggtt tatcaaagtg gcgcatgatg tccctctcac 780agtcacgaag tagcaaatct gcaccagaag gagggacaat tatctatatg ccacctgaaa 840actatgaacc tggacaaaaa tcaagggcca gtatcaagca cgatatatat agctatgcag 900ttatcacatg ggaagtgtta tccagaaaac agccttttga agatgtcacc aatcctttgc 960agataatgta tagtgtgtca caaggacatc gacctgttat taatgaagaa agtttgccat 1020atgatatacc tcaccgagca cgtatgatct ctctaataga aagtggatgg gcacaaaatc 1080cagatgaaag accatctttc ttaaaatgtt taatagaact tgaaccagtt ttgagaacat 1140ttgaagagat aacttttctt gaagctgtta ttcagctaaa gaaaacaaag ttacagagtg 1200tttcaagtgc cattcaccta tgtgacaaga agaaaatgga attatctctg aacatacctg 1260taaatcatgg tccacaagag gaatcatgtg gatcctctca gctccatgaa aatagtggtt 1320ctcctgaaac ttcaaggtcc ctgccagctc ctcaagacaa tgatttttta tctagaaaag 1380ctcaagactg ttattttatg aagctgcatc actgtcctgg aaatcacagt tgggatagca 1440ccatttctgg atctcaaagg gctgcattct gtgatcacaa gaccactcca tgctcttcag 1500caataataaa tccactctca actgcaggaa actcagaacg tctgcagcct ggtatagccc 1560agcagtggat ccagagcaaa agggaagaca ttgtgaacca aatgacagaa gcctgcctta 1620accagtcgct agatgccctt ctgtccaggg acttgatcat gaaagaggac tatgaacttg 1680ttagtaccaa gcctacaagg acctcaaaag tcagacaatt actagacact actgacatcc 1740aaggagaaga atttgccaaa gttatagtac aaaaattgaa agataacaaa caaatgggtc 1800ttcagcctta cccggaaata cttgtggttt ctagatcacc atctttaaat ttacttcaaa 1860ataaaagcat gtaagtgact gtttttcaag aagaaatgtg tttcataaaa ggatatttat 1920atctctgttg ctttgacttt ttttatataa aatccgtgag tattaaagct ttattgaagg 1980ttctttgggt aaatattagt ctccctccat gacactgcag tatttttttt aattaataca 2040agtaaaaagt ttgaattttg ctacatagtt caatttttat gtctcttttg ttaacagaaa 2100ccacttttaa aggatagtaa ttattcttgt ttataacagt gccttaaggt atgatgtatt 2160tctgatggaa gccattttca cattcatgtt cttcatggat tatttgttac ttgtctaaga 2220tgcaatttga ttttatgaag tatataccct ttacccacca gagacagtac agaatccctg 2280ccctaaaatc ccaggcttaa ttgccctaca aagggttatt aatttaaaac tccattatta 2340ggattacatt ttaaagtttt atttatgaat tccctttaaa aatgatattt caaaggtaaa 2400acaatacaat ataaagaaaa aaataaatat attaataccg gcttcctgtc cccattttta 2460acctcagcct tccctactgt caccaacaac caagctaaat aaagtcaaca gcctgatgtg 2520t 252125537DNAHomo sapiens 2gaacccggcg aggaaataca tgcactggct gagaatcgcc cgcgccaggg cgcaacgcca 60caaggtgtag ggagtgtgcg gggtggggcg aaaggggacc caagagtccc tgtggctcgg 120agtgccgggc cgtcggttct tcattcctgc cctcggggca gacggagtga ccccggcccc 180cactccccgc cccgaccatg gtagtgttca atggccttct taagatcaaa atctgcgagg 240ccgtgagctt gaagcccaca gcctggtcgc tgcgccatgc ggtgggaccc cggccgcaga 300ctttccttct cgacccctac attgccctca atgtggacga ctcgcgcatc ggccaaacgg 360ccaccaagca gaagaccaac agcccggcct ggcacgacga gttcgtcacc gatgtgtgca 420acggacgcaa gatcgagctg gctgtctttc acgatgcccc cataggctac gacgacttcg 480tggccaactg caccatccag tttgaggagc tgctgcagaa cgggagccgc cacttcgagg 540actggattga tctggagcca gaaggaagag tgtatgtgat catcgatctc tcagggtcgt 600cgggtgaagc ccctaaagac aatgaagagc gtgtgttcag ggaacgcatg cggccgagga 660agcggcaggg ggccgtcagg cgcagggtcc atcaggtcaa cggccacaag ttcatggcca 720cctatcttcg gcagcccacc tactgctccc attgcagaga cttcatctgg ggtgtcatag 780gaaagcaggg ataccagtgt caagtctgca cctgcgtggt ccacaagcgg tgccacgagc 840tcataatcac aaagtgtgct gggttaaaga agcaggagac ccccgaccag gtgggctccc 900agcggttcag cgtcaacatg ccccacaagt tcggtatcca caactacaag gtccctacct 960tctgcgatca ctgtgggtcc ctgctctggg gactcttgcg gcagggtttg cagtgtaaag 1020tctgcaaaat gaatgttcac cgtcgatgtg agaccaacgt ggctcccaac tgtggagtgg 1080atgccagagg aatcgccaaa gtactggccg acctgggcgt taccccagac aaaatcacca 1140acagcggcca gagaaggaaa aagctcattg ctggtgccga gtccccgcag cctgcttctg 1200gaagctcacc atctgaggaa gatcgatcca agtcagcacc cacctcccct tgtgaccagg 1260aaataaaaga acttgagaac aacattcgga aagccttgtc atttgacaac cgaggagagg 1320agcaccgggc agcatcgtct cctgatggcc agctgatgag ccccggtgag aatggcgaag 1380tccggcaagg ccaggccaag cgcctgggcc tggatgagtt caacttcatc aaggtgttgg 1440gcaaaggcag ctttggcaag gtcatgttgg cagaactcaa gggcaaagat gaagtatatg 1500ctgtgaaggt cttaaagaag gacgtcatcc ttcaggatga tgacgtggac tgcacaatga 1560cagagaagag gattttggct ctggcacgga aacacccgta ccttacccaa ctctactgct 1620gcttccagac caaggaccgc ctctttttcg tcatggaata tgtaaatggt ggagacctca 1680tgtttcagat tcagcgctcc cgaaaattcg acgagcctcg ttcacggttc tatgctgcag 1740aggtcacatc ggccctcatg ttcctccacc agcatggagt catctacagg gatttgaaac 1800tggacaacat ccttctggat gcagaaggtc actgcaagct ggctgacttc gggatgtgca 1860aggaagggat tctgaatggt gtgacgacca ccacgttctg tgggactcct gactacatag 1920ctcctgagat cctgcaggag ttggagtatg gcccctccgt ggactggtgg gccctggggg 1980tgctgatgta cgagatgatg gctggacagc ctccctttga ggccgacaat gaggacgacc 2040tatttgagtc catcctccat gacgacgtgc tgtacccagt ctggctcagc aaggaggctg 2100tcagcatctt gaaagctttc atgacgaaga atccccacaa gcgcctgggc tgtgtggcat 2160cgcagaatgg cgaggacgcc atcaagcagc acccattctt caaagagatt gactgggtgc 2220tcctggagca gaagaagatc aagccaccct tcaaaccacg cattaaaacc aaaagagacg 2280tcaataattt tgaccaagac tttacccggg aagagccggt actcaccctt gtggacgaag 2340caattgtaaa gcagatcaac caggaggaat tcaaaggttt ctcctacttt ggtgaagacc 2400tgatgccctg agagcccact gcagttggac tttgccgatg ctgcaagaag gggtgcagag 2460aagactcctg tgttggagac actcagcagg tcttgaacta cttctcctcc tcggagcccc 2520agtcccatgt ccactgtcta tttattgcat tcccttgccc caggccacct cctccccctc 2580ccacctggtg accagaaggc gctctcggtt cttgtctcac cagtaatgca gactcattgg 2640gtcagcaatt agctgtatac actgccgtgt ttggaccatt ggcaagcctg gttccactcc 2700tcaggggctc ctggcagtga agcaacttca gttcttttac tgcaaagaac agaaaaaaga 2760aagaaagcaa acaagaagac tccggctctg ctatcggaca cagatcctga tccctcttgc 2820ttcttttccc tcctgcaccg cagcttgcca tccctgccct tctgtcctgg agaagagact 2880ggtgcttctc cgcacacacg agggagggcg cccttgaggc atgccctctg agggagggag 2940accagagatg cagggattgg ccagctgggt tggtttgctc tggaatggct aactcttgcc 3000tgctttggtt ttagcttttc agcatgccaa agtcatgtaa gtttgtgtct tgtggaagaa 3060atcctctttg tggaaaaaga aacagggttt tgaactctgt taacatttga aaaatatatt 3120ttcaaattca ctttctaatt ggccaaaaga gatgagttcc agtctgaata caggtagata 3180ttaaagggct aataaaaaat gagaaaccgg tcgtccaagg tggatgctgt caatgcccga 3240gtgacacatg agagctgtat gaattgagag aaaaggcaac aagtagcatt cttcatcatt 3300caagttctac ctggacacaa aggcgaggac cctggggttc caacaaagct cagctcccag 3360attctctttc cagtttcatc ctaagttcct agcataaaca ctatttattt tctgcagcag 3420tgtgttattt ttgcgcactt atacaaaatg gtagtactac tgtgttgtgg tttttaaaca 3480ttaaacatgt aaagttatat acgaaatatc tgcttttgga ataagcagaa tgaggctaaa 3540catgggttat acaaagggta tctggaaact gaagagcaac ttgttagaaa actgacaatg 3600tcgcaagatg tactcagttt tgtttctgtg tgacatgcaa tggcaactca tgtggacact 3660attgaaggga tgtgacatta cctcctgtag atatgctaac agtgttattc tttcatttcc 3720aagggttctc tgtggctttg tgtatatgtt tcccagaggt catttgatta cctaatttac 3780tgaactgatt tagcagggaa tggaatccat tccaactatt gcacgtggat ttcccagctg 3840cccctaaata tatatacttg tgagtggcaa agtggcacta atgaagcttt tgccttttgt 3900acatttgaga tttttgtata tagtgtttgc tgcaaggcct gtggaattaa ttcgttgcat 3960atagaggtat caactgctgc atgttcaggc atattataaa actttagtct atgaaagaat 4020aattataata atgtccaggt gcaatactct gtaagtctat tggttcaagt taccgagaga 4080taggtgtgtt cctttatggg ggatgggggg gtgtgttggg gattctttgt attgtttatt 4140tcattttggt ttattttaaa agatgtaaac atatattaag ctatattaaa tctcacatac 4200agttcttctg tgctctatta taccctgata gagatggggg agagaaagga atgtttttga 4260tggtggtttc aaagctcgga cagtaactat cttgagccca ttagagagtc tgtgtccata 4320tttgcatctg gctggtcata gcctttgtta ctaatgatga cattcagttc tcttttgttt 4380ttatttttta aaaactcagg tgtaattatt atctgttctt aagataattg caaatattaa 4440atattatgat atatcaattc atgtgtttgg cataccagtg aatgatgaag aacatgagat 4500taatttaatt tatcttcggt aacttgacat tctggagaga gactatcttc tggagttgag 4560tacaagcaca gaaacatctt tacggtggca tcatctcatt ttttaggaag acatgataat 4620actgcccatc atattcatgt gtaactactg ttctttcttc tgctttcttc accataataa 4680actttggaca accaagcaag ctctaaccgc aatgccagat ggccttgtcc gagggcctag 4740tgtttgcacg gcagtgggaa ctgggccttt cctacaggac aactggcaag tttgctggga 4800agtcaaataa tacattccac ctggcagctg aaggcagcca gtcagtctgt cccagaaagg 4860gcccttttca gcacccaaag ctgggctggc tgggatgcct ctggctggtg aagttctcac 4920ataggctgat ttaaatccag caaaggtcta tagaaaaagg cttgcgtgtt cgttgagtaa 4980tcattgtttc attttcattt ttacgagagt ttgaaaatag acacactgtt aacacttctg 5040ccagtttttt ctgatctttc cagccccacc ccctttctct ttctctctct ctctcaaaga 5100aaaaaaaaat gggagtgcaa aaaaaacaaa gccaaaaaat atatgaagga tagctgttct 5160tctgtgttct ctcattatgg actttgtgaa gtagaaacat aatttttttt cctccaaagg 5220tgaaaaaaca atgcattctt gctttaaaaa aaaaaaagaa ggctaaaaaa ttacctcttt 5280ttaaattatg tgcaaaataa ttctggctaa ctgtaaaatg tattcaattt taggattttt 5340tttttttgta ttgtgatgct ttatttgtac atttttttcc tttctggatg taattttaat 5400ctcttgccat tcattagtgt tatttcattg taaacgttat tgtgccaaat gtactgtatt 5460caaaaggatg tgaatgtgta ttgtttcaga acctaataaa tacaatgacg ttaagtctta 5520aaaaaaaaaa aaaaaaa 553731969DNAHomo sapiens 3gcccgcgggc ctcgccgccc cgcgcggatc gtcgcggccc ggccgtcccg tcccaggaag 60tggccgtcct gagcgccatg gctcactccc cggtgcagtc gggcctgccc ggcatgcaga 120acctaaaggc agacccagaa gagcttttta caaaactaga gaaaattggg aagggctcct 180ttggagaggt gttcaaaggc attgacaatc ggactcagaa agtggttgcc ataaagatca 240ttgatctgga agaagctgaa gatgagatag aggacattca acaagaaatc acagtgctga 300gtcagtgtga cagtccatat gtaaccaaat attatggatc ctatctgaag gatacaaaat 360tatggataat aatggaatat cttggtggag gctccgcact agatctatta gaacctggcc 420cattagatga aacccagatc gctactatat taagagaaat actgaaagga ctcgattatc 480tccattcgga gaagaaaatc cacagagaca ttaaagcggc caacgtcctg ctgtctgagc 540atggcgaggt gaagctggcg gactttggcg tggctggcca gctgacagac acccagatca 600aaaggaacac cttcgtgggc accccattct ggatggcacc cgaggtcatc aaacagtcgg 660cctatgactc gaaggcagac atctggtccc tgggcataac agctattgaa cttgcaagag 720gggaaccacc tcattccgag ctgcacccca tgaaagtttt attcctcatt ccaaagaaca 780acccaccgac gttggaagga aactacagta aaccsctcaa ggagtttgtg gaggcctgtt 840tgaataagga gccgagcttt agacccactg ctaaggagtt attgaagcac aagtttatac 900tacgcaatgc aaagaaaact tcctacttga ccgagctcat cgacaggtac aagagatgga 960aggccgagca gagccatgac gactcgagct ccgaggattc cgacgcggaa acagatggcc 1020aagcctcggg gggcagtgat tctggggact ggatcttcac aatccgagaa aaagatccca 1080agaatctcga gaatggagct cttcagccat cggacttgga cagaaataag atgaaagaca 1140tcccaaagag gcctttctct cagtgtttat ctacaattat ttctcctctg tttgcagagt 1200tgaaggagaa gagccaggcg tgcggaggga acttgggttc cattgaagag ctgcgagggg 1260ccatctacct agcggaggag gtgtgccctg gcatctccga caccatggtg gcccagctcg 1320tgcagcggct ccagagatac tctctaagtg gtggaggaac ttcatcccac tgaaattcct 1380ttggcatttg gggttttgtt tttccttttt tccttcttca tcctcctcct tttttaaaag 1440tcaacgagag ccttcgctga ctccaccgaa gaggtgcgcc actgggagcc accccagcsc 1500caggcgcccg tccagggaca cacacagtct tcgctgtgct gcagccagat gaagtctctc 1560agatgggtgg ggagggtcag ctccttccag cgatcatttt attttatttt attacktttg 1620tttttaattt taaccatagc gcacatattc caggaaagtg tctttaaaaa caaaaacaaa 1680ccctgaaatg tatatttggg attatgataa ggcaactaaa gacatgaaac ctcaggtatc 1740ctgctttaag ttgataactc cctctgggag ctggagaatc gctctggtgg atgggtgtac 1800agatttgtat ataatgtcat ttttacggaa accctttcgg cgtgcataag gaatcactgt 1860gtacaaactg gccaagtgct tctgtagata acgtcagtgg agtaaatatt cgacaggcca 1920taacttgagt ctattgcctt gcctttatta catgtacatt ttgaattcc 196942505DNAHomo sapiens 4tttgggttag ggagagtgct ttcgtttgtt ttaaatggga gaaactggag catgttgcca 60aggcagagag ccagcagaga ggggtgaatg gaagaaggag cgagaagggg gttactgacg 120aagccttatc ctggaggaga gaaggatgga ctccagagcc cagctttggg gactggcctt 180gaataaaagg agggccactc tacctcatcc tggagggagc acgaacctaa aggcagaccc 240agaagagctt tttacaaaac tagagaaaat tgggaagggc tcctttggag aggtgttcaa 300aggcattgac aatcggactc agaaagtggt tgccataaag atcattgatc tggaagaagc 360tgaagatgag atagaggaca ttcaacaaga aatcacagtg ctgagtcagt gtgacagtcc 420atatgtaacc aaatattatg gatcctatct gaaggataca aaattatgga taataatgga 480atatcttggt ggaggctccg cactagatct attagaacct ggcccattag atgaaaccca 540gatcgctact atattaagag aaatactgaa aggactcgat tatctccatt cggagaagaa 600aatccacaga gacattaaag cggccaacgt cctgctgtct gagcatggcg aggtgaagct 660ggcggacttt ggcgtggctg gccagctgac agacacccag atcaaaagga acaccttcgt 720gggcacccca ttctggatgg cacccgaggt catcaaacag tcggcctatg actcgaaggc 780agacatctgg tccctgggca taacagctat tgaacttgca agaggggaac cacctcattc 840cgagctgcac cccatgaaag ttttattcct cattccaaag aacaacccac cgacgttgga 900aggaaactac agtaaacccc tcaaggagtt tgtggaggcc tgtttgaata aggagccgag 960ctttagaccc actgctaagg agttattgaa gcacaagttt atactacgca atgcaaagaa 1020aacttcctac ttgaccgagc tcatcgacag gtacaagaga tggaaggccg agcagagcca 1080tgacgactcg agctccgagg attccgacgc ggaaacagat ggccaagcct cggggggcag 1140tgattctggg gactggatct tcacaatccg agaaaaagat cccaagaatc tcgagaatgg 1200agctcttcag ccatcggact tggacagaaa taagatgaaa gacatcccaa agaggccttt 1260ctctcagtgt ttatctacaa ttatttctcc tctgtttgca gagttgaagg agaagagcca 1320ggcgtgcgga gggaacttgg ggtccattga agagctgcga ggggccatct acctagcgga 1380ggaggcgtgc cctggcatct ccgacaccat ggtggcccag ctcgtgcagc ggctccagag 1440atactctcta agtggtggag gaacttcatc ccactgaaat tcctttggca tttggggttt 1500tgtttttcct tttttccttc ttcatcctcc tcctttttta aaagtcaacg agagccttcg 1560ctgactccac cgaagaggtg cgccactggg agccacccca gcgccaggcg cccgtccagg 1620gacacacaca gtcttcactg tgctgcagcc agatgaagtc tctcagatgg gtggggaggg 1680tcagctcctt ccagcgatca ttttatttta ttttattact tttgttttta attttaacca 1740tagtgcacat attccaggaa agtgtcttta aaaacaaaaa caaaccctga aatgtatatt 1800tgggattatg ataaggcaac taaagacatg aaacctcagg tatcctgctt taagttgata 1860actccctctg gagcttggag aatcgctctg gtggatgggt gtacagattt gtatataatg 1920tcatttttac ggaaaccctt tcggcgtgca taaggaatca ctgtgtacaa actggccaag 1980tgcttctgta gataacgtca gtggagtaaa tattcgacag gccataaact tgagtctatt 2040gccttgcctt tattacatgt acattttgaa ttctgtgacc agtgatttgg gttttatttt 2100gtatttgcag ggtttgtcat taataattaa tgcccctctc ttacagaaca ctcctatttg 2160tacctcaaca aatgcaaatt ttccccgttt gccctacgcc ccttttggta cacctagagg 2220ttgatttcct ttttcatcga tggtactatt tcttagtgtt ttaaattgga acatatcttg 2280cctcatgaag ctttaaatta taattttcag tttctcccca tgaagcgctc tcgtctgaca 2340tttgtttgga atcgtgccac tgctggtctg cgccagatgt accgtccttt ccaatacgat 2400tttctgttgc accttgtagt ggattctgca tatcatcttt cccacctaaa aatgtctgaa 2460tgcttacaca aataaatttt ataacacgct taaaaaaaaa aaaaa 250552060DNAHomo sapiens 5gcggccgcgt ggggcccagc acaaagacct gtccccaggg gccgccgcct ccgccgctgc 60tgctgccgcc agcctagagc cgcccgccga agcagagccg gcgccggggt cctcatcccc 120accggtcccg aggggcggct gctgcccgtc gccacgaggc ccaggggccc gagtgccgag 180ccctttgctc cctcggccgc gcggggacag ggctgctgag cagcctccgc ctctcccggc 240tgtgggggcc ccactgagta tgtcggagga gagcgacatg gacaaagcca tcaaggaaac 300ttccatttta gaagaataca gtatcaattg gactcagaag ctgggagctg gaattagtgg 360tccagttaga gtctgtgtaa agaaatctac tcaagaacgg tttgcgctga aaattcttct 420tgatcgtcca aaagctagaa atgaggtacg tctgcacatg atgtgtgcca cacacccaaa 480catagttcag attattgaag tgtttgctaa cagtgtccag tttccccatg agtccagccc 540tagggcccga ctcttaattg taatggagat gatggaaggg ggagagctat ttcacagaat 600cagccagcac cggcacttta cagagaagca agccagccaa gtaacaaagc agatagcttt 660ggctctgcgg cactgtcact tgttaaacat tgcgcacaga gacctcaagc ctgaaaatct 720gctttttaag gataactctt tggatgcccc agtgaagttg tgtgactttg gatttgccaa 780gattgaccaa ggtgacttga tgacacccca gttcacccct tattatgtag caccccaggt 840actggaggcg caaagaaggc atcagaagga gaaatctggc atcataccta cctcaccgac 900gccctacact tacaacaaga gctgtgactt gtggtcccta ggggtgatta tctatgtgat 960gctgtgcgga taccctcctt tttactccaa acaccacagc cggactatcc caaaggatat 1020gcgaagaaag atcatgacag gcagttttga gttcccagag gaagagtgga gtcagatctc 1080agagatggcc aaagatgttg tgaggaagct cctgaaggtc aaaccggagg agagactcac 1140catcgaggga gtgctggacc acccctggct caattccacc gaggccctgg ataatgtgct 1200gccttctgct cagctgatga tggacaaggc agtggttgca ggaatccagc aggctcacgc 1260ggaacagttg gccaacatga gaatccagga tctgaaagtc agcctcaaac ccctgcactc 1320agtgaacaac cccattctgc ggaagaggaa gttacttggc accaagccaa aggacagtgt 1380ctatatccac gaccatgaga atggagccga ggattccaat gttgccttgg aaaaactccg 1440agatgtgatt gctcagtgta ttctccccca ggctggagag aatgaagatg agaaactgaa 1500tgaagtaatg caggaggctt ggaagtataa ccgggaatgc aaactcctaa gagatactct 1560gcagagcttc agctggaatg gtcgtggatt cacagataaa gtagatcgac taaaactggc 1620agaaattgtg aagcaggtga tagaagagca aaccacgtcc cacgaatccc aataatgaca 1680gcttcagact ttgttttttt aacaatttga aaaattattc tttaatgtat aaagtaattt 1740tatgtaaatt aataaatcat aatttcattt ccacattgat taaagctgct gtatagattt 1800agggtgcagg acttaataat agtatagtta ttgtttgttt ttaagaaaag ctcagttcta 1860gagacatact attactttag gactgtgtag ttgtatattt gtaagatgac agatgatgct 1920gtcaagcaat attgttttat ttgtaataaa atatacaaaa atcacttgcc agcagtagaa 1980aaaggaccga ctataccgac ctttctgatt agtaaacagt tgaatcaagg actctggaaa 2040aaaaaaaaaa aaaaaaaaaa 206062066DNAHomo sapiens 6gcggccgcgt ggggcccagc acaaagacct gtccccaggg gccgccgcct ccgccgctgc 60tgctgccgcc agcctagagc cgcccgccga agcagagccg gcgccggggt cctcatcccc 120accggtcccg aggggcggct gctgcccgtc gccacgaggc ccaggggccc gagtgccgag
180ccctttgctc cctcggccgc gcggggacag ggctgctgag cagcctccgc ctctcccggc 240tgtgggggcc ccactgagta tgtcggagga gagcgacatg gacaaagcca tcaaggaaac 300ttccatttta gaagaataca gtatcaattg gactcagaag ctgggagctg gaattagtgg 360tccagttaga gtctgtgtaa agaaatctac tcaagaacgg tttgcgctga aaattcttct 420tgatcgtcca aaagctagaa atgaggtacg tctgcacatg atgtgtgcca cacacccaaa 480catagttcag attattgaag tgtttgctaa cagtgtccag tttccccatg agtccagccc 540tagggcccga ctcttaattg taatggagat gatggaaggg ggagagctat ttcacagaat 600cagccagcac cggcacttta cagagaagca agccagccaa gtaacaaagc agatagcttt 660ggctctgcgg cactgtcact tgttaaacat tgcgcacaga gacctcaagc ctgaaaatct 720gctttttaag gataactctt tggatgcccc agtgaagttg tgtgactttg gatttgccaa 780gattgaccaa ggtgacttga tgacacccca gttcacccct tattatgtag caccccaggt 840actggaggcg caaagaaggc atcagaagga gaaatctggc atcataccta cctcaccgac 900gccctacact tacaacaaga gctgtgactt gtggtcccta ggggtgatta tctatgtgat 960gctgtgcgga taccctcctt tttactccaa acaccacagc cggactatcc caaaggatat 1020gcgaagaaag atcatgacag gcagttttga gttcccagag gaagagtgga gtcagatctc 1080agagatggcc aaagatgttg tgaggaagct cctgaaggtc aaaccggagg agagactcac 1140catcgaggga gtgctggacc acccctggct caattccacc gaggccctgg ataatgtgct 1200gccttctgct cagctgatga tggacaaggc agtggttgca ggaatccagc aggctcacgc 1260ggaacagttg gccaacatga gaatccagga tctgaaagtc agcctcaaac ccctgcactc 1320agtgaacaac cccattctgc ggaagaggaa gttacttggc accaagccaa aggacagtgt 1380ctatatccac gaccatgaga atggagccga ggattccaat gttgccttgg aaaaactccg 1440agatgtgatt gctcagtgta ttctccccca ggctggtaaa ggagagaatg aagatgagaa 1500actgaatgaa gtaatgcagg aggcttggaa gtataaccgg gaatgcaaac tcctaagaga 1560tactctgcag agcttcagct ggaatggtcg tggattcaca gataaagtag atcgactaaa 1620actggcagaa attgtgaagc aggtgataga agagcaaacc acgtcccacg aatcccaata 1680atgacagctt cagactttgt ttttttaaca atttgaaaaa ttattcttta atgtataaag 1740taattttatg taaattaata aatcataatt tcatttccac attgattaaa gctgctgtat 1800agatttaggg tgcaggactt aataatagta tagttattgt ttgtttttaa gaaaagctca 1860gttctagaga catactatta ctttaggact gtgtagttgt atatttgtaa gatgacagat 1920gatgctgtca agcaatattg ttttatttgt aataaaatat acaaaaatca cttgccagca 1980gtagaaaaag gaccgactat accgaccttt ctgattagta aacagttgaa tcaaggactc 2040tggaaaaaaa aaaaaaaaaa aaaaaa 206672736DNAHomo sapiens 7gcgaccgctc cccggcggga gccagcgaag gtttccatgt cagaggccga tggagaactg 60aagattgcca cctacgcaca aaggccattg agacacttcg tgtagctgga agacaccaac 120ttcctgacag gagctttatt tcatttggga tttcaagttt acagatggta tcttctcaaa 180agttggaaaa acctatagag atgggcagta gcgaacccct tcccatcgca gatggtgaca 240ggaggaggaa gaagaagcgg aggggccggg ccactgactc cttgccagga aagtttgaag 300atatgtacaa gctgacctct gaattgcttg gagagggagc ctatgccaaa gttcaaggtg 360ccgtgagcct acagaatggc aaagagtatg ccgtcaaaat catcgagaaa caagcagggc 420acagtcggag tagggtgttt cgagaggtgg agacgctgta tcagtgtcag ggaaacaaga 480acattttgga gctgattgag ttctttgaag atgacacaag gttttacttg gtctttgaga 540aattgcaagg aggttccatc ttagcccaca tccagaagca aaagcacttc aatgagcgag 600aagccagccg agtggtgcgg gacgttgctg ctgcccttga cttcctgcat accaaagaca 660aagtctctct ctgtcaccta ggctggagtg ctatggcgcc atcagggctc actgcagccc 720caacctccct gggctccagt gatcctccca cctcagcctc ccaagtagct gggactacag 780gcattgctca tcgtgatctg aaaccagaaa atatattgtg tgaatctcca gaaaaggtgt 840ctccagtgaa aatctgtgac tttgacttgg gcagtgggat gaaactgaac aactcctgta 900cccccataac cacaccagag ctgaccaccc catgtggctc tgcagaatac atggcccctg 960aggtagtgga ggtcttcacg gaccaggcca cattctacga caagcgctgt gacctgtgga 1020gcctgggcgt ggtcctctac atcatgctga gtggctaccc acccttcgtg ggtcactgcg 1080gggccgactg tggctgggac cggggcgagg tctgcagggt gtgccagaac aagctgtttg 1140aaagcatcca ggaaggcaag tatgagtttc ctgacaagga ctgggcacac atctccagtg 1200aagccaaaga cctcatctcc aagctcctgg tgcgagatgc aaagcagaga cttagcgccg 1260cccaagttct gcagcaccca tgggtgcagg ggcaagctcc agaaaaggga ctccccacgc 1320cgcaagtcct ccagaggaac agcagcacaa tggacctgac gctcttcgca gctgaggcca 1380tcgcccttaa ccgccagcta tctcagcacg aagagaacga actagcagag gagccagagg 1440cactagctga tggcctctgc tccatgaagc tttcccctcc ctgcaagtca cgcctggccc 1500ggagacgggc cctggcccag gcaggccgtg gtgaagacag gagcccgccc acagcactct 1560gaaatgctcc agtcacacct tataggccct aggcctggcc aggcattgtc ccctggaaac 1620ctgtgtggct aaagtctgct gagcaggcag cagcctctgc tctgtggctc cattcaggct 1680ttttcatcta cgaaggccct gaggttccca tcaaccccca tttccctagg gtcctggagg 1740aaaaagcttt ttccaaaggg gttgtctttg aaaaggaaag caatcacttc tcactttgca 1800taattgcctg cagcaggaac atctcttcac tgggctccac ctgctcaccc gcctgcagat 1860ctgggatcca gcctgctctc accgctgtag ctgtggcggc tggggctgca gcctgcaggg 1920agaagcaaga agcatcagtt gacagaggct gccgacacgt gcctcttccc tctcttctct 1980gtcaccctcc tctggcggtc cttccacctt cctctgtcct ccggatgtcc tctttgcccg 2040tcttctccct tggctgagca aagccatccc ctcaattcag ggaagggcaa ggagccttcc 2100tcattcagga aatcaaatca gtcttccggt ctgcagcacg gaaaagcaca taatctttct 2160ttgctgtgac tgaaatgtat ccctcgttta tcatcccctt tgtttgtgat tgctgctaaa 2220gtcagtagta tcgttttttt aaaaaaaaag tttggtgttt ttaaccatgc tgttccagca 2280aagatgatac cttaaactcc cactgcaagc ccatgaactt cccagagagt ggaacggctt 2340gctcttcttt ctagaatgtc catgcacttg ggttttaatc agcagttccc tattattctg 2400attttaagct gttcctgtga tgaacttaga gacagcatcg gtgtctgctg ctgtgtcccc 2460aggtcttgtg tgggtggcac agatctgggc agttagatag tgctctgtgc ctaaggtgaa 2520gccacactag ggtgaagcct cacttccctg tttgagcaat gcagtgcctg ctgcccgtgt 2580gcatgaaggt acagccattc agataagtgg aactattgag ttacataaag aaaatagatt 2640tgcatttgtc aggcagacgt ttatacaaca ccacggtgct tttatacatt gtgcttattt 2700taataaaact gaaattctaa aaaaaaaaaa aaaaaa 273682168DNAHomo sapiens 8ctctctcgct cctgcgttcg caggcggcgg ctggcggccg gcttctcgct cgggcagcgg 60cggcggcggc ggcggcggct tccggagtcc cgctgcgaag atgctcaaag tcacggtgcc 120ctcctgctcc gcctcgtcct gctcttcggt caccgccagt gcggccccgg ggaccgcgag 180cctcgtcccg gattactgga tcgacggctc caacagggat gcgctgagcg atttcttcga 240ggtggagtcg gagctgggac ggggtgctac atccattgtg tacagatgca aacagaaggg 300gacccagaag ccttatgctc tcaaagtgtt aaagaaaaca gtggacaaaa aaatcgtaag 360aactgagata ggagttcttc ttcgcctctc acatccaaac attataaaac ttaaagagat 420atttgaaacc cctacagaaa tcagtctggt cctagaactc gtcacaggag gagaactgtt 480tgataggatt gtggaaaagg gatattacag tgagcgagat gctgcagatg ccgttaaaca 540aatcctggag gcagttgctt atctacatga aaatgggatt gtccatcgtg atctcaaacc 600agagaatctt ctttatgcaa ctccagcccc agatgcacca ctcaaaatcg ctgattttgg 660actctctaaa attgtggaac atcaagtgct catgaagaca gtatgtggaa ccccagggta 720ctgcgcacct gaaattctta gaggttgtgc ctatggacct gaggtggaca tgtggtctgt 780aggaataatc acctacatct tactttgtgg atttgaacca ttctatgatg aaagaggcga 840tcagttcatg ttcaggagaa ttctgaattg tgaatattac tttatctccc cctggtggga 900tgaagtatct ctaaatgcca aggacttggt cagaaaatta attgttttgg atccaaagaa 960acggctgact acatttcaag ctctccagca tccgtgggtc acaggtaaag cagccaattt 1020tgtacacatg gataccgctc aaaagaagct ccaagaattc aatgcccggc gtaagcttaa 1080ggcagcggtg aaggctgtgg tggcctcttc gcgcctggga agtgccagca gcagccatgg 1140cagcatccag gagagccaca aggctagccg agacccttct ccaatccaag atggcaacga 1200ggacatgaaa gctattccag aaggagagaa aattcaaggc gatggggccc aagccgcagt 1260taagggggca caggctgagc tgatgaaggt gcaagcctta gagaaagtta aaggtgcaga 1320tataaatgct gaagaggccc ccaaaatggt gcccaaggca gtggaggatg ggataaaggt 1380ggctgacctg gaactagagg agggcctagc agaggagaag ctgaagactg tggaggaggc 1440agcagctccc agagaagggc aaggaagctc tgctgtgggt tttgaagttc cacagcaaga 1500tgtgatcctg ccagagtact aaacagcttc cttcagatct ggaagccaaa caccggcatt 1560ttatgtactt tgtccttcag caagaaaggt gtggaagcat gatatgtact atagtgattc 1620tgtttttgag gtgcaaaaaa catacatata taccagttgg taattctaac ttcaatgcat 1680gtgactgctt tatgaaaata atagtgtctt ctatggcatg taatggatac ctaataccga 1740tgagttaaat cttgcaagtt aacacaacgt aacacttaaa agcatacatt ttcagcaacc 1800agtggcacat atttgaagtg aatagtagca aattgttttt gctttgaaaa tctagccatc 1860ctacatcctt tggatttctt cacaaggcag taattccttt gaactactgc ttagctaata 1920ctaggtagtg ctaaaagaca tgttcccata acttttacaa cattttactt tttatcattg 1980atgtgttcaa actgtttaca aggagatgct tatagatgat agttgtacat atgtgcaaaa 2040aaaaatccac ttgcaatggt aagaaattga agtatcctta aaggccatga agccatatgt 2100ccctaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 2160aaaaaaaa 216891634DNAHomo sapiens 9ccctgctaac aaagggagcc acttccttcc tctctgcaca taccccatgt ctcaccacga 60tgatggagct acagtgggac ttggaatcca gatgtgtgaa ggatggaggg ttgaagccgc 120actcagcttc ctgccccacc agaggaagtg gggagagacg gcaggtgcag tgatggctgg 180cggagtcatg gacaaggagt acgtgggttt tgctgccctc cccaaccagc tgcaccgcaa 240gtctgtcaag aaggggtttg acttcacgct aatggtggca ggggagtcag gcctagggaa 300atccaccctc atcaacagcc tcttcctcac caacctctat gaggatcgcc aggtgccaga 360ggccagtgct cgcttgacac agaccctggc cattgagcgc cggggcgtag agattgagga 420agggggtgtg aaagtgaagc tgacccttgt ggacacacct ggctttgggg actcagtgga 480ctgctctgac tgctggcttc cggtggtgaa attcatcgag gagcaatttg agcagtacct 540tagggatgag agtggcctga accggaagaa catccaggac tcccgagtcc actgctgcct 600ctacttcatc tcacccttcg gccgggggct ccggccccta gatgtggcct tcctccgggc 660agtacacgag aaagtcaaca tcatcccagt cattggcaaa gcggatgctc tgatgcccca 720ggaaacccag gccctcaagc agaagatccg ggatcagttg aaggaagagg agatccacat 780ctaccagttc cccgaatgtg actctgatga agatgaagac ttcaagaggc aggatgcaga 840gatgaaggaa agcatccctt ttgcagtcgt gggatcatgc gaggtggtga gggatggcgg 900gaaccggccg gtgaggggac gccgctactc ctgggggacc gtggaggtgg agaacccaca 960tcactgcgat ttcctgaacc tgcgacggat gctggtgcag acacacctgc aggacctgaa 1020agaggtgacg cacgatctgc tctacgaggg ctaccgggcc cgctgcctac agagcctggc 1080ccggcctggg gctcgcgatc gagccagccg cagtaagctt tcccgccaga gcgccacaga 1140gatcccgctg cccatgctgc ctctggcgga caccgagaag ctgatccgcg agaaagacga 1200agagctgcgc cgcatgcaag agatgctgga gaagatgcag gcccaaatgc agcagagcca 1260ggcccagggc gagcagtcag acgccctctg aggccacgcc ccgcccggcc ttacctcggc 1320tccgccttca gtcggcctct tgtccaatcc ccgcgcccca cactgcccag cgccccccgg 1380gacctccgcg ggtgccgccc tcgcgcgggc tagggggagg ttctcccagc ctgagtccgt 1440agccccgccc cggcgctggt cccgcccacc cagacaccgc ccacttcccg gcccggggcc 1500tgcacaatct ccgaccgcat cactgtcttc cggagtcccc cttcttctcc cagactctgt 1560cttcaataaa aactgagctt cccgcggcca aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 1620aaaaaaaaaa aaaa 163410747DNAHomo sapiens 10ccgagatgtg cagcggcaca gctgtcgcgc cagtcgcaac agaagcaggt ccgaggcaca 60gcccgatccc gccatggagc agccgaggaa ggcggtggta gtgacgggat ttggcccttt 120tggggaacac accgtgaacg ccagttggat tgcagttcag gagctagaaa agctaggcct 180tggcgacagc gtggacctgc atgtgtacga gattccggtt gagtaccaaa cagtccagag 240actcatcccc gccctgtggg agaagcacag tccacagctg gtggtgcatg tgggggtgtc 300aggcatggcg accacagtca cactggagaa atgtggacac aacaagggct acaaggggct 360ggacaactgc cgcttttgcc ccggctccca gtgctgcgtg gaggacgggc ctgaaagcat 420tgactccatc atcgacatgg atgctgtgtg caagcgagtc accacgttgg gcctggatgt 480gtcggtgacc atctcgcagg atgccggcag gaaaaaaccc ttccctgcca aaggtgactg 540tgttttctgc cgccgaagga gggcccggtc cctccaggct cagtgtggct tctccctgac 600ccccgcccta gaacttttgc cagtgccttt tctgaaactc ctgtgtcccg ggccccccag 660gcggagaagg atatgccgga ttctgcctgg ggctgggctc taggagaccc caaatttgac 720accacagaac gcacatacta cacttga 747112239DNAHomo sapiens 11agtcgcaaca gaagcaggtc cgaggcacag cccgatcccg ccatggagca gccgaggaag 60gcggtggtag tgacgggatt tggccctttt ggggaacaca ccgtgaacgc cagttggatt 120gcagttcagg agctagaaaa gctaggcctt ggcgacagcg tggacctgca tgtgtacgag 180attccggttg agtaccaaac agtccagaga ctcatccccg ccctgtggga gaagcacagt 240ccacagctgg tggtgcatgt gggggtgtca ggcatggcga ccacagtcac actggagaaa 300tgtggacaca acaagggcta caaggggctg gacaactgcc gcttttgccc cggctcccag 360tgctgcgtgg aggacgggcc tgaaagcatt gactccatca tcgacatgga tgctgtgtgc 420aagcgagtca ccacgttggg cctggatgtg tcggtgacca tctcgcagga tgccggcaga 480tatctctgcg actttaccta ctacacctct ttgtaccaga gtcacggtcg atcagccttc 540gtccacgtgc ccccactggg gaagccgtac aacgcggacc agctgggcag ggcactgaga 600gccatcattg aggagatgtt ggacctcctg gagcagtcag agggcaaaat caactattgc 660cacaaacact gagggacgct caggtctcct aagacctcat cctgctgggg accccacgag 720gggacatcca ccctctgggg tgtggccagg aaaagacaag ctcttcagct tggggatccg 780atctggaaga gagattctga tctgcccacc tcctcttcct tcttctctac aaaagctccg 840gttgattcga gggaagtggt gaaaattttt ttttctccca ttttcctccc tgcatctggg 900gacacagctg ccgtgaccag ggaggccagc ctgggaggtc cagatgccca gggagaatct 960tggtctggtg aatccatgag ctgagatacc acggctgggg ccatatgttc acctgctttc 1020ctgtccgttg gtgaaggaat ttcagaattc attttatatc caagactggc ttttaccaaa 1080tttaaaagcc tctcaatgcg tcctcgacct tgaactgtgc tcaacagcct ggccctttct 1140ggggccaccc tgggatatgg ctggctggct ggctggcttt ctttctttct ttctttcttt 1200ctttctttct ttctttcttt ctttctttct ttcttgcttt ctttcttgct ttctttcttg 1260ctttctttct ttcttgcttt ctttcttctc tctctctctt tttttttttt ttaaatagtg 1320ctagtttggg cacagagtaa tttatattcc ctttggttaa aatgcaggct ttttagccaa 1380caacaaaagt gttttccccc cacccccact cgcccaccag ggtgatgcca cttttgcctc 1440ctgccctgaa aattggactt aagatgccat gtcttggctg ggattacagg catgagccac 1500tgcacccagc ccagttcttt cttaaaacag ctgagagttt tgttttcttc agcgttcacc 1560ttccttgtct ccagttccga tgctggcagt ggttcctacc tctgttgggt ttctagatag 1620tttgggaacg gggttgatgg gtttctgtga aacacatttt ccaagtcttg ggctttctct 1680ggaggggaag gtggatgctg gcgggtgact tgcagtgggc gcctggcagt gggtgtggac 1740tgtaactgac aggtggaaat gagtaggggc actattgttc cctccatgcc agcttttttt 1800ttgctggaaa tgccccctcc acacccctgg tagctctgtg tcctgagaaa tccagagtgt 1860gggagacatc actgcatctg tccccccagc ttctgtgaag ggaagctgtg gcctcttttg 1920aatgtgggga acaactgaag actcaggggt cacccagagg tctggtggaa agcaacttca 1980ggtttcatct tgctctattc ctcaaaggtc tggtctgtgg gcctctgagg agaaaacagg 2040tctagccaag acagggacaa aatggggaag ggggtgtgcc aggcctgaac tgagctaagc 2100acctgccccg ggctccacac ttccatcttt cttttgtctt catttcacct ctgtgtttaa 2160agcactgtgt gacatagctc cttagagata taacctattg tctgctcatt gtcaaaaaaa 2220aaaaaaaaaa aaaaaaaaa 2239121531DNAHomo sapiens 12agtcacagag ggaacacaga gcctagttgt aaacggacag agacgagagg ggcaagggag 60gacagtggat gacagggaag acgagtgggg gcagagctgc tcaggaccat ggctgaggcc 120atcacctatg cagatctgag gtttgtgaag gctcccctga agaagagcat ctccagccgg 180ttaggacagg acccaggggc tgatgatgat ggggaaatca cctacgagaa tgttcaagtg 240cccgcagtcc taggggtgcc ctcaagcttg gcttcttctg tactagggga caaagcagcg 300gtcaagtcgg agcagccaac tgcgtcctgg agagccgtga cgtcaccagc tgtcgggcgg 360attctcccct gccgcacaac ctgcctgcga tacctcctgc tcggcctgct cctcacctgc 420ctgctgttag gagtgaccgc catctgcctg ggagtgcgct atctgcaggt gtctcagcag 480ctccagcaga cgaacagggt tctggaagtc actaacagca gcctgaggca gcagctccgc 540ctcaagataa cgcagctggg acagagtgca gaggatctgc aggggtccag gagagagctg 600gcgcagagtc aggaagcact acaggtggaa cagagggctc atcaggcggc cgaagggcag 660ctacaggcct gccaggcaga cagacagaag acgaaggaga ccttgcaaag tgaggagcaa 720cagaggaggg ccttggagca gaagctgagc aacatggaga acagactgaa gcccttcttc 780acatgcggct cagcagacac ctgctgtccg tcgggatgga taatgcatca gaaaagctgc 840ttttacatct cacttacttc aaaaaattgg caggagagcc aaaaacaatg tgaaactctg 900tcttccaagc tggccacatt cagtgaaatt tatccacaat cacactctta ctacttctta 960aattcactgt tgccaaatgg tggttcaggg aattcatatt ggactggcct cagctctaac 1020aaggattgga agttgactga tgatacacaa cgcactagga cttatgctca aagctcaaaa 1080tgtaacaagg tacataaaac ttggtcatgg tggacactgg agtcagagtc atgtagaagt 1140tctcttccct acatctgtga gatgacagct ttcaggtttc cagattagga cagtcctttg 1200cactgagttg acactcatgc caacaagaac ctgtgcccct ccttcctaac ctgaggcctg 1260gggttcctca gaccatctcc ttcattctgg gcagtgccag ccaccggctg acccacacct 1320gacacttcca gccagtctgc tgcctgctcc ctcttcctga aactggactg ttcctgggaa 1380aagggtgaag ccacctctag aagggacttt ggcctccccc caagaacttc ccatggtaga 1440atggggtggg ggaggagggc gcacgggctg agcggatagg ggcggcccgg agccagccag 1500gcagttttat tgaaatcttt ttaaataatt g 1531132033DNAHomo sapiens 13gactagttct agatcgcgat ctagaactag ccgcgggaga cgctgctgag gcggcttcgg 60ttgcgggtcg gaacggcgct gctctgcggg gccggtccag gctggcagct gccggcgctt 120ggcggtgagg gcgggctccc gagtggcccc ccaccgaagg cggtgggacc agcggctgag 180gccaggatgc cgtccaggcg gcgcggcggc tcctcactca tcccagatgt tggttatctt 240tctgaagtag actgtccatg gcctgaacat tttccgaaaa tcattttgag caaaatatct 300gtttaataac aagataacca catcaagatg gttggaaagc tgaagcagaa cttactattg 360gcatgtctgg tgattagttc tgtgactgtg ttttacctgg gccagcatgc catggaatgc 420catcaccgga tagaggaacg tagccagcca gtcaaattgg agagcacaag gaccactgtg 480agaactggcc tggacctcaa agccaacaaa acctttgcct atcacaaaga tatgccttta 540atatttattg gaggtgtgcc tcggagtgga accacactca tgagggccat gctggacgca 600catcctgaca ttcgctgtgg agaggaaacc agggtcattc cccgaatcct ggccctgaag 660cagatgtggt cacggtcaag taaagagaag atccgcctgg atgaggctgg tgttactgat 720gaagtgctgg attctgccat gcaagccttc ttactagaaa ttatcgttaa gcatggggag 780ccagcccctt atttatgtaa taaagatcct tttgccctga aatctttaac ttacctttct 840aggttattcc ccaatgccaa atttctcctg atggtccgag atggccgggc atcagtacat 900tcaatgattt ctcgaaaagt tactatagct ggatttgatc tgaacagcta tagggactgt 960ttgacaaagt ggaatcgtgc tatagagacc atgtataacc agtgtatgga ggttggttat 1020aaaaagtgca tgttggttca ctatgaacaa cttgtcttac atcctgaacg gtggatgaga 1080acactcttaa agttcctcca gattccatgg aaccactcag tattgcacca tgaagagatg 1140attgggaaag ctgggggagt gtctctgtca aaagtggaga gatctacaga ccaagtaatc 1200aagccagtca atgtaggagc tctatcaaaa tgggttggga agataccgcc agatgtttta 1260caagacatgg cagtgattgc tcctatgctt gccaagcttg gatatgaccc atatgccaac 1320ccacctaact acggaaaacc tgatcccaaa attattgaaa acactcgaag ggtctataag 1380ggagaattcc aactacctga ctttcttaaa gaaaaaccac agactgagca agtggagtag 1440cagaaccagg agcctcttcc atacatgagg aaagattgct gccttttcag cagaagggaa 1500attcctagga ttggctgtcc cctgccaagc ttggtggagc gtctgcacct tggctgcgcc 1560gcctgtgcat ttgccagttt cctcccactg agaggatgga ggtgtccgca cagctttggg 1620cctcgtgagg gatctgcctc ctgagcaaag agctcttgat cccgatttca tgcacagccc 1680tgcagtaagg agcccagaag gaacatgtgt ttcctgttaa
aactcctctt gttctctttt 1740cttacattat gacgtttgtt ttcaaggaga gggtttaaaa atgggatcct gtaagcagac 1800ttgggcagtc tccttttgaa ataggttgtc tgtacatgtt ctaatgtttt gtagaacacg 1860tgtgcctgtt taagtgtatt gatgtgaata atattaaata tcctaattat ttaattcatt 1920gtattgtttc tgagaagttg ggaaattacc attatacatt tacaacctaa tgacttttgt 1980attttatttt tcaaaataaa agctttcaat gtgaagcaaa aaaaaaaaaa aaa 2033141050DNAHomo sapiens 14atgaactcca ccttggatgg taatcagagc agccaccctt tttgcctctt ggcatttggc 60tatttggaaa ctgtcaattt ttgccttttg gaagtattga ttattgtctt tctaactgta 120ttgattattt ctggcaacat cattgtgatt tttgtatttc actgtgcacc tttgttgaac 180catcacacta caagttattt tatccagact atggcatatg ctgacctttt tgttggggtg 240agctgcgtgg tcccttcttt atcactcctc catcaccccc ttccagtaga ggagtccttg 300acttgccaga tatttggttt tgtagtatca gttctgaaga gcgtctccat ggcttctctg 360gcctgtatca gcattgatag atacattgcc attactaaac ctttaaccta taatactctg 420gttacaccct ggagactacg cctgtgtatt ttcctgattt ggctatactc gaccctggtc 480ttcctgcctt cctttttcca ctggggcaaa cctggatatc atggagatgt gtttcagtgg 540tgtgcggagt cctggcacac cgactcctac ttcaccctgt tcatcgtgat gatgttatat 600gccccagcag cccttattgt ctgcttcacc tatttcaaca tcttccgcat ctgccaacag 660cacacaaagg atatcagcga aaggcaagcc cgcttcagca gccagagtgg ggagactggg 720gaagtgcagg cctgtcctga taagcgctat gccatggtcc tgtttcgaat cactagtgta 780ttttacatcc tctggttgcc atatatcatc tacttcttgt tggaaagctc cactggccac 840agcaaccgct tcgcatcctt cttgaccacc tggcttgcta ttagtaacag tttctgcaac 900tgtgtaattt atagtctctc caacagtgta ttccaaagag gactaaagcg cctctcaggg 960gctatgtgta cttcttgtgc aagtcagact acagccaacg acccttacac agttagaagc 1020aaaggccctc ttaatggatg tcatatctga 1050152190DNAHomo sapiens 15ggtgaagccg gtggccggtg gccgggcggg accaacaaag atggcggcgg cccctgcggc 60gggagcgatc tgggcaacgg ctgcggctaa agctgcagcc gggcccacgg gggggctgca 120cgggggtagt agggggtggc cctgaactgg ggcctggccc tggctggcct ctcccgccgc 180ctcactgggg gacaggtcca gcctgtggtg tccacaatgc cccaggcctc tgagcaccgc 240ctgggccgta cccgagagcc acctgttaat atccagcccc gagtgggatc caagctacca 300tttgccccca gggcccgcag caaggagcgc agaaacccag cctctgggcc aaaccccatg 360ttacgacctc tgcctccccg gccaggtctg cctgatgaac ggctcaagaa actggagctg 420ggacggggac ggacctcagg ccctcgtccc agaggccccc ttcgagcaga tcatggggtt 480cccctgcctg gctcaccacc cccaacagtg gctttgcctc tcccatctcg gaccaactta 540gcccgttcca agtctgtgag cagtggggac ttgcgtccaa tggggattgc cttgggaggg 600caccgtggca ccggagagct tggggctgca ctgagccgct tggccctccg gcctgagcca 660cccactttga gacgtagcac ttctctccgc cgcctagggg gctttcctgg accccctacc 720ctgttcagca tacggacaga gccccctgct tcccatggct ccttccacat gatatccgcc 780cggtcctctg agcctttcta ctctgatgac aagatggctc atcacacact ccttctgggc 840tctggtcatg ttggccttcg aaacctggga aacacgtgct tcctgaatgc tgtgctgcag 900tgtctgagca gcactcgacc tcttcgggac ttctgtctga gaagggactt ccggcaagag 960gtgcctggag gaggccgagc ccaagagctc actgaagcct ttgcagatgt gattggtgcc 1020ctctggcacc ctgactcctg cgaagctgtg aatcctactc gattccgagc tgtcttccag 1080aaatatgttc cctccttctc tggatacagc cagcaggatg cccaagagtt cctgaagctc 1140ctcatggagc ggctacacct tgaaatcaac cgccgaggcc gccgggctcc accgatactt 1200gccaatggtc cagttccctc tccaccccgc cgaggagggg ctctgctaga agaacctgag 1260ttaagtgatg atgaccgagc caacctaatg tggaaacgtt acctggagcg agaggacagc 1320aagattgtgg acctgtttgt gggccagttg aaaagttgtc tcaagtgcca ggcctgtggg 1380tatcgctcca cgaccttcga ggttttttgt gacctgtccc tgcccatccc caagaaagga 1440tttgctgggg gcaaggtgtc tctgcgggat tgtttcaacc ttttcactaa ggaagaagag 1500ctagagtcgg agaatgcccc agtgtgtgac cgatgtcggc agaaaactcg aagtaccaaa 1560aagttgacag tacaaagatt ccctcgaatc ctcggcttag atctgaatcg attttctgcc 1620tcccgaggct ccatcaaaaa aagttcagta ggtgtagact ttccactgca gcgactgagc 1680ctaggggact ttgccagtga caaagccgga agtcctgtat accagctgta tgccctttgc 1740aaccactcag gcagcgtcca ctatggccac tacacagccc tgtgccggtg ccagactggt 1800tggcatgtct acaatgactc tcgtgtctcc cctgtcagtg aaaaccaggt ggcatccagc 1860gagggctacg tgctgttcta ccaactgatg caggagccac cccggtgcct gtgacacctc 1920taagctctgg cacctgtgaa gccctttaaa cacccttaag ccccaggctc cccgtttacc 1980tcagagacgt ctatttttgt gtctttttaa tcggggaggg gggagggggt ggttgtagct 2040ccattatttt ttttattaaa aaataccctt ccacctggag gctcccttgt ctcccagccc 2100catgtacaaa gctcaccaag cccctgccca tgtacagccc ccagaccctc tgcaatatca 2160ctttttgtga ataaatttat taagaaaaaa 2190162148DNAHomo sapiens 16ggtgaagccg gtggccggtg gccgggcggg accaacaaag atggcggcgg cccctgcggc 60gggagcgatc tgggcaacgg ctgcggctaa agctgcagcc gggcccacgg gggggctgca 120cgggggtagt agggggtggc cctgaactgg ggcctggccc tggctggcct ctcccgccgc 180ctcactgggg gacaggtcca gcctgtggtg tccacaatgc cccaggcctc tgagcaccgc 240ctgggccgta cccgagagcc acctgttaat atccagcccc gagtgggatc caagctacca 300tttgccccca gggcccgcag caaggagcgc agaaacccag cctctgggcc aaaccccatg 360ttacgacctc tgcctccccg gccaggtctg cctgatgaac ggctcaagaa actggagctg 420ggacggggac ggacctcagg ccctcgtccc agaggccccc ttcgagcaga tcatggggtt 480cccctgcctg gctcaccacc cccaacagtg gctttgcctc tcccatctcg gaccaactta 540gcccgttcca agtctgtgag cagtggggac ttgcgtccaa tggggattgc cttgggaggg 600caccgtggca ccggagagct tggggctgca ctgagccgct tggccctccg gcctgagcca 660cccactttga gacgtagcac ttctctccgc cgcctagggg gctttcctgg accccctacc 720ctgttcagca tacggacaga gccccctgct tcccatggct ccttccacat gatatccgcc 780cggtcctctg agcctttcta ctctgatgac aagatggctc atcacacact ccttctgggc 840tctggtcatg ttggccttcg aaacctggga aacacgtgct tcctgaatgc tgtgctgcag 900tgtctgagca gcactcgacc tcttcgggac ttctgtctga gaagggactt ccggcaagag 960gtgcctggag gaggccgagc ccaagagctc actgaagcct ttgcagatgt gattggtgcc 1020ctctggcacc ctgactcctg cgaagctgtg aatcctactc gattccgagc tgtcttccag 1080aaatatgttc cctccttctc tggatacagc cagcaggatg cccaagagtt cctgaagctc 1140ctcatggagc ggctacacct tgaaatcaac cgccgaggcc gccgggctcc accgatactt 1200gccaatggtc cagttccctc tccaccccgc cgaggagggg ctctgctaga agaacctgag 1260ttaagtgatg atgaccgagc caacctaatg tggaaacgtt acctggagcg agaggacagc 1320aagattgtgg acctgtttgt gggccagttg aaaagttgtc tcaagtgcca ggcctgtggg 1380tatcgctcca cgaccttcga ggttttttgt gacctgtccc tgcccatccc caagaaagga 1440tttgctgggg gcaaggtgtc tctgcgggat tgtttcaacc ttttcactaa ggaagaagag 1500ctagagtcgg agaatgcccc agtgtgtgac cgatgtcggc agaaaactcg aagtaccaaa 1560aagttgacag tacaaagatt ccctcgaatc ctcggcttag atctgaatcg attttctgcc 1620tcccgaggct ccatcaaaaa aagttcagta ggtgtagact ttccactgca gcgactgagc 1680ctaggggact ttgccagtga caaagccggc agcgtccact atggccacta cacagccctg 1740tgccggtgcc agactggttg gcatgtctac aatgactctc gtgtctcccc tgtcagtgaa 1800aaccaggtgg catccagcga gggctacgtg ctgttctacc aactgatgca ggagccaccc 1860cggtgcctgt gacacctcta agctctggca cctgtgaagc cctttaaaca cccttaagcc 1920ccaggctccc cgtttacctc agagacgtct atttttgtgt ctttttaatc ggggaggggg 1980gagggggtgg ttgtagctcc attatttttt ttattaaaaa atacccttcc acctggaggc 2040tcccttgtct cccagcccca tgtacaaagc tcaccaagcc cctgcccatg tacagccccc 2100agaccctctg caatatcact ttttgtgaat aaatttatta agaaaaaa 2148177391DNAHomo sapiens 17gctgcgcagc gctggctgct ggctggcctc gcggagacgc cgaacggacg cggccggcgc 60cggcttgtgg gctcgccgcc tgcagccatg accctcgcag cctgtccctc ggcctcggcc 120cgggacgtct aaaatcccac acagtcgcgc gcagctgctg gagagccggc cgctgccccc 180tcgtcgccgc atcacactcc cgtcccggga gctgggagca gcgcgggcag ccggcgcccc 240cgtgcaaact gggggtgtct gccagagcag ccccagccgc tgccgctgct acccccgatg 300ctggccatgg cctggcgggg cgcagggccg agcgtcccgg gggcgcccgg gggcgtcggt 360ctcagtctgg ggttgctcct gcagttgctg ctgctcctgg ggccggcgcg gggcttcggg 420gacgaggaag agcggcgctg cgaccccatc cgcatctcca tgtgccagaa cctcggctac 480aacgtgacca agatgcccaa cctggttggg cacgagctgc agacggacgc cgagctgcag 540ctgacaactt tcacaccgct catccagtac ggctgctcca gccagctgca gttcttcctt 600tgttctgttt atgtgccaat gtgcacagag aagatcaaca tccccattgg cccatgcggc 660ggcatgtgtc tttcagtcaa gagacgctgt gaacccgtcc tgaaggaatt tggatttgcc 720tggccagaga gtctgaactg cagcaaattc ccaccacaga acgaccacaa ccacatgtgc 780atggaagggc caggtgatga agaggtgccc ttacctcaca aaacccccat ccagcctggg 840gaagagtgtc actctgtggg aaccaattct gatcagtaca tctgggtgaa aaggagcctg 900aactgtgtgc tcaagtgtgg ctatgatgct ggcttataca gccgctcagc caaggagttc 960actgatatct ggatggctgt gtgggccagc ctgtgtttca tctccactgc cttcacagta 1020ctgaccttcc tgatcgattc ttctaggttt tcctaccctg agcgccccat catatttctc 1080agtatgtgct ataatattta tagcattgct tatattgtca ggctgactgt aggccgggaa 1140aggatatcct gtgattttga agaggcagca gaacctgttc tcatccaaga aggacttaag 1200aacacaggat gtgcaataat tttcttgctg atgtactttt ttggaatggc cagctccatt 1260tggtgggtta ttctgacact cacttggttt ttggcagcag gactcaaatg gggtcatgaa 1320gccattgaaa tgcacagctc ttatttccac attgcagcct gggccatccc cgcagtgaaa 1380accattgtca tcttgattat gagactggtg gatgcagatg aactgactgg cttgtgctat 1440gttggaaacc aaaatctcga tgccctcacc gggttcgtgg tggctcccct ctttacttat 1500ttggtcattg gaactttgtt cattgctgca ggtttggtgg ccttgttcaa aattcggtca 1560aatcttcaaa aggatgggac aaagacagac aagttagaaa gactgatggt caagattggg 1620gtgttctcag tactgtacac agttcctgca acgtgtgtga ttgcctgtta tttttatgaa 1680atctccaact gggcactttt tcggtattct gcagatgatt ccaacatggc tgttgaaatg 1740ttgaaaattt ttatgtcttt gttggtgggc atcacttcag gcatgtggat ttggtctgcc 1800aaaactcttc acacgtggca gaagtgttcc aacagattgg tgaattctgg aaaggtaaag 1860agagagaaga gaggaaatgg ttgggtgaag cctggaaaag gcagtgagac tgtggtataa 1920ggctagtcag cctccatgct ttcttcattt tgaagggggg aatgccagca ttttggagga 1980aattctacta aaagttttat gcagtgaatc tcagtttgaa caaactagca acaattaagt 2040gacccccgtc aacccactgc ctcccacccc gaccccagca tcaaaaaacc aatgattttg 2100ctgcagactt tggaatgatc caaaatggaa aagccagtta gaggctttca aagctgtgaa 2160aaatcaaaac gttgatcact ttagcaggtt gcagcttgga gcgtggaggt cctgcctaga 2220ttccaggaag tccagggcga tactgttttc ccctgcaggg tgggatttga gctgtgagtt 2280ggtaactagc agggagaaat attaactttt ttaacccttt accattttaa atactaactg 2340ggtctttcag atagcaaagc aatctataaa cactggaaac gctgggttca gaaaagtgtt 2400acaagagttt tatagtttgg ctgatgtaac ataaacatct tctgtggtgc gctgtctgct 2460gtttagaact ttgtggactg cactcccaag aagtggtgtt agaatctttc agtgcctttg 2520tcataaaaca gttatttgaa caaacaaaag tactgtactc acacacataa ggtatccagt 2580ggatttttct tctctgtctt cctctcttaa atttcaacat ctctcttctt ggctgctgct 2640gttttcttca ttttatgtta atgactcaaa aaaggtattt ttatagaatt tttgtactgc 2700agcatgctta aagaggggaa aaggaagggt gattcacttt ctgacaatca cttaattcag 2760aggaaaatga gatttactaa gttgacttac ctgacggacc ccagagacct attgcattga 2820gcagtgggga cttaatatat tttacttgtg tgattgcatc tatgcagacg ccagtctgga 2880agagctgaaa tgttaagttt cttggcaact ttgcattcac acagattagc tgtgtaattt 2940ttgtgtgtca attacaatta aaagcacatt gttggaccat gacatagtat actcaactga 3000ctttaaaact atggtcaact tcaacttgca ttctcagaat gatagtgcct ttaaaaattt 3060ttttattttt taaagcataa gaatgttatc agaatctggt ctacttagga caatggagac 3120tttttcagtt ttataaaggg aactgaggac agctaatcca actacttggt gcgtaattgt 3180ttcctagtaa ttggcaaagg ctccttgtaa gatttcactg gaggcagtgt ggcctggagt 3240atttatatgg tgcttaatga atctccagaa tgccagccag aagcctgatt ggttagtagg 3300gaataaagtg tagaccatat gaaatgaact gcaaactcta atagcccagg tcttaattgc 3360ctttagcaga ggtatccaaa gcttttaaaa tttatgcata cgttcttcac aagggggtac 3420ccccagcagc ctctcgaaaa ttgcacttct cttaaaactg taactggcct ttctcttacc 3480ttgccttagg ccttctaatc atgagatctt ggggacaaat tgactatgtc acaggttgct 3540ctccttgtaa ctcatacctg tctgcttcag caactgcttt gcaatgacat ttatttatta 3600attcatgcct taaaaaaata ggaagggaag cttttttttt tctttttttt tttttcaatc 3660acactttgtg gaaaaacatt tccagggact caaaattcca aaaaggtggt caaattctgg 3720aagtaagcat ttcctctttt ttaaaaattt ggtttgagcc ttatgcccat agtttgacat 3780ttccctttct tctttccttt ttgtttttgt gtggttcttg agctctctga catcaagatg 3840catgtaaagt cgattgtatg ttttggaagg caaagtcttg gcttttgaga ctgaagttaa 3900gtgggcacag gtggcccctg ctgctgtgcc cagtctgagt accttggcta gactctaggt 3960caggctccag gagcatgaga attgatcccc agaagaacca ttttaactcc atctgatact 4020ccattgccta tgaaatgtaa aatgtgaact ccctgtgctg cttgtagaca gttcccataa 4080ctgtccacgg ccctggagca cgcacccagg ggcagagcct gcccttactc acgctctgct 4140ctggtgtctt gggagttgtg cagggactct ggcccaggca ggggaaggaa gaccaggcgg 4200taggggactg gtcttgctgt tagagtatag aggtttgtaa tgcagttttc ttcataatgt 4260gtcagtgatt gtgtgaccaa ggcagcatct agcagaaagc caggcatgga gtaggtgatc 4320gatacttgtc aatgactaaa taataacaat aaaagagcac ttgggtgaat ctgggcacct 4380gatttctgag ttttgagttc tggagctagt gttttgacaa tgctttgggt tttgacatgc 4440cttttccaca aatctcttgc cttttcaggg caaagtgtat ttgatcagaa gtggccattt 4500ggattagtag ccttagcaat gctacagggt tataggcctc tcctttcaca ttccagacaa 4560tggagagtgt ttatggtttc aggaaaagaa ctttgtggct gaggggtcag ttaccagtga 4620ccttcaatca actccatcac ttcttaaatc ggtatttgtt aaaaaaatca gttattttat 4680ttattgagtg ccgactgtag taaagccctg aaatagataa tctctgttct tctaactgat 4740ctaggatggg gacgcaccca ggtctgctga actttactgt tcctctggga aaggagcagg 4800gacctctgga attcccatct gtttcactgt ctccattcca taaatctctt cctgtgtgag 4860ccaccacacc cagcctgggt ctctctactt ttaacacatc tctcatccct ttcccaggat 4920tccttccaag tcagttacag gtggttttaa cagaaagcat cagctctgct tcgtgacagt 4980ctctggagaa atcccttagg aagactatga gagtaggcca caaggacatg ggcccacaca 5040tctgctttgg ctttgccggc aattcagggc ttggggtatt ccatgtgact tgtataggta 5100tatttgagga cagcatcttg ctagagaaaa ggtgagggtt gtttttcttt ctctgaaacc 5160tacagtaaat gggtatgatt gtagcttcct cagaaatccc ttggcctcca gagattaaac 5220atggtgcaat ggcacctctg tccaacctcc tttctggtag attcctttct cctgcttcat 5280ataggccaaa cctcagggca agggaacatg ggggtagagt ggtgctggcc agaaccatct 5340gcttgagcta cttggttgat tcatatcctc tttcctttat ggagacccat ttcctgatct 5400ctgagactgt tgctgaactg gcaacttact tgggcctgaa actggagaag gggtgacatt 5460tttttaattt cagagatgct ttctgatttt cctctcccag gtcactgtct cacctgcact 5520ctccaaactc aggttccggg aagcttgtgt gtctagatac tgaattgaga ttctgttcag 5580caccttttag ctctatactc tctggctccc ctcatcctca tggtcactga attaaatgct 5640tattgtattg agaaccaaga tgggacctga ggacacaaag atgagctcaa cagtctcagc 5700cctagaggaa tagactcagg gatttcacca ggtcggtgca gtatttgatt tctggtgagg 5760tgaccacagc tgcagttagg gaagggagcc attgagcaca gactttggaa ggaacctttt 5820ttttgttgtt tgtttgtttg tttgtttgtt tgtttgtttg agacagggtc ttgctctgtc 5880acccaggctg gggcgcaatg gcacgatctt ggctcactgc aacctctgcc tcctgggttc 5940aagtgattct cctgccacag cctcctgagg agctgggact acaggtgcgt gctaccacgc 6000ccagctactt ctgtattttt agtagagacg gggtttcact gtgttggcca ggctggtctc 6060gaactcctga cctcatgatc tgcccgcctc agcctcccaa agtgctggga ttacaagtgt 6120gagccaccac acctggcctg gaaggaacct cttaaaatca gtttacgtct tgtattttgt 6180tctgtgatgg aggacactgg agagagttgc tattccagtc aatcatgtcg agtcactgga 6240ctctgaaaat cctattggtt cctttatttt atttgagttt agagttccct tctgggtttg 6300tattatgtct ggcaaatgac ctgggttatc acttttcctc cagggttaga tcatagatct 6360tggaaactcc ttagagagca ttttgctcct accaaggatc agatactgga gccccacata 6420atagatttca tttcactcta gcctacatag agctttctgt tgctgtctct tgccatgcac 6480ttgtgcggtg attacacact tgacagtacc aggagacaaa tgacttacag atcccccgac 6540atgcctcttc cccttggcaa gctcagttgc cctgatagta gcatgtttct gtttctgatg 6600tacctttttt ctcttcttct ttgcatcagc caattcccag aatttcccca ggcaatttgt 6660agaggacctt tttggggtcc tatatgagcc atgtcctcaa agcttttaaa cctccttgct 6720ctcctacaat attcagtaca tgaccactgt catcctagaa ggcttctgaa aagaggggca 6780agagccactc tgcgccacaa aggttgggtc catcttctct ccgaggttgt gaaagttttc 6840aaattgtact aataggctgg ggccctgact tggctgtggg ctttgggagg ggtaagctgc 6900tttctagatc tctcccagtg aggcatggag gtgtttctga attttgtcta cctcacaggg 6960atgttgtgag gcttgaaaag gtcaaaaaat gatggcccct tgagctcttt gtaagaaagg 7020tagatgaaat atcggatgta atctgaaaaa aagataaaat gtgacttccc ctgctctgtg 7080cagcagtcgg gctggatgct ctgtggcctt tcttgggtcc tcatgccacc ccacagctcc 7140aggaaccttg aagccaatct gggggacttt cagatgtttg acaaagaggt accaggcaaa 7200cttcctgcta cacatgccct gaatgaattg ctaaatttca aaggaaatgg accctgcttt 7260taaggatgta caaaagtatg tctgcatcga tgtctgtact gtaaatttct aatttatcac 7320tgtacaaaga aaaccccttg ctatttaatt ttgtattaaa ggaaaataaa gttttgtttg 7380ttaaaaaaaa a 7391181302DNAHomo sapiens 18atgtgcacag agaagatcaa catccccatt ggcccatgcg gcggcatgtg tctttcagtc 60aagagacgct gtgaacccgt cctgaaggaa tttggatttg cctggccaga gagtctgaac 120tgcagcaaat tcccaccaca gaacgaccac aaccacatgt gcatggaagg gccaggtgat 180gaagaggtgc ccttacctca caaaaccccc atccagcctg gggaagagtg tcactctgtg 240ggaaccaatt ctgatcagta catctgggtg aaaaggagcc tgaactgtgt gctcaagtgt 300ggctatgatg ctggcttata cagccgctca gccaaggagt tcactgatat ctggatggct 360gtgtgggcca gcctgtgttt catctccact gccttcacag tactgacctt cctgatcgat 420tcttctaggt tttcctaccc tgagcgcccc atcatatttc tcagtatgtg ctataatatt 480tatagcattg cttatattgt caggctgact gtaggccggg aaaggatatc ctgtgatttt 540gaagaggcag cagaacctgt tctcatccaa gaaggactta agaacacagg atgtgcaata 600attttcttgc tgatgtactt ttttggaatg gccagctcca tttggtgggt tattctgaca 660ctcacttggt ttttggcagc aggactcaaa tggggtcatg aagccattga aatgcacagc 720tcttatttcc acattgcagc ctgggccatc cccgcagtga aaaccattgt catcttgatt 780atgagactgg tggatgcaga tgaactgact ggcttgtgct atgttggaaa ccaaaatctc 840gatgccctca ccgggttcgt ggtggctccc ctctttactt atttggtcat tggaactttg 900ttcattgctg caggtttggt ggccttgttc aaaattcggt caaatcttca aaaggatggg 960acaaagacag acaagttaga aagactgatg gtcaagattg gggtgttctc agtactgtac 1020acagttcctg caacgtgtgt gattgcctgt tatttttatg aaatctccaa ctgggcactt 1080tttcggtatt ctgcagatga ttccaacatg gctgttgaaa tgttgaaaat ttttatgtct 1140ttgttggtgg gcatcacttc aggcatgtgg atttggtctg ccaaaactct tcacacgtgg 1200cagaagtgtt ccaacagatt ggtgaattct ggaaaggtaa agagagagaa gagaggaaat 1260ggttgggtga agcctggaaa aggcagtgag actgtggtat aa 1302191998DNAHomo sapiens 19cggcgcgatg cgcggagacc cccgcggggg cggcggcggc cgtgagcccc gatgaggccc 60gagcgtcccc ggccgcgcgg cagcgccccc ggcccgatgg agaccccgcc gtgggaccca 120gcccgcaacg actcgctgcc gcccacgctg accccggccg tgccccccta cgtgaagctt 180ggcctcaccg tcgtctacac cgtgttctac gcgctgctct tcgtgttcat ctacgtgcag 240ctctggctgg tgctgcgtta ccgccacaag cggctcagct accagagcgt cttcctcttt 300ctctgcctct tctgggcctc cctgcggacc gtcctcttct ccttctactt caaagacttc 360gtggcggcca attcgctcag ccccttcgtc ttctggctgc
tctactgctt ccctgtgtgc 420ctgcagtttt tcaccctcac gctgatgaac ttgtacttca cgcaggtgat tttcaaagcc 480aagtcaaaat attctccaga attactcaaa taccggttgc ccctctacct ggcctccctc 540ttcatcagcc ttgttttcct gttggtgaat ttaacctgtg ctgtgctggt aaagacggga 600aattgggaga ggaaggttat cgtctctgtg cgagtggcca ttaatgacac gctcttcgtg 660ctgtgtgccg tctctctctc catctgtctc tacaaaatct ctaagatgtc cttagccaac 720atttacttgg agtccaaggg ctcctccgtg tgtcaagtga ctgccatcgg tgtcaccgtg 780atactgcttt acacctctcg ggcctgctac aacctgttca tcctgtcatt ttctcagaac 840aagagcgtcc attcctttga ttatgactgg tacaatgtat cagaccaggc agatttgaag 900aatcagctgg gagatgctgg atacgtatta tttggagtgg tgttatttgt ttgggaactc 960ttacctacca ccttagtcgt ttatttcttc cgagttagaa atcctacaaa ggaccttacc 1020aaccctggaa tggtccccag ccatggattc agtcccagat cttatttctt tgacaaccct 1080cgaagatatg acagtgatga tgaccttgcc tggaacattg cccctcaggg acttcaggga 1140ggttttgctc cagattacta tgattgggga caacaaacta acagcttcct ggcacaagca 1200ggaactttgc aagactcaac tttggatcct gacaaaccaa gccttgggta gcatcagtta 1260acagttttat ggacgattcc tcagatgaaa agcttcagaa aagcatagtg acagctgaat 1320ttttagggca cttttcctta agaaatagaa cttgattttt atttgttaca ggtttccaat 1380ggccccatag gaataagcaa taatgtagac tgataaaccc ttattttagt actaaagagg 1440gagccttgct atttcagtgg gtataattta aactttttaa agaaaatctg tacttttata 1500aagatgtatt ttgtataact taaataataa tgctaaagta tactagggtt tttttttctt 1560gagaatgtta ctgcaatcat gttgtagttt gcacagactt ttatgcataa ttcactttaa 1620aaatatagaa tatatggtct aatagttttt taaagctttt ggactaaagt attccacaaa 1680tcttacctct ttaggtcact gatggtcact ccgattctga gtgccacatt ggtagactcc 1740taaaatacag ttgacaactt agccaattgc aactccagtg ttgataatta aaatgaaatg 1800gtaaagcagc agactgtaag gtctttagag attttttttt aaggttcagg ccgtaggttc 1860ctcaaggaat ctcttaagtt ttgcccaaag actggtactt cctttcagta gggcgctaat 1920gtatacacat taatgataag ttgataacat taaaaatgta gctgacttat cctattaaac 1980ctcctctgct atgttcac 199820890DNAHomo sapiens 20ctggccacac cctccgcctg gacgcagcag ccaccgccgc gtccctctct ccacgaggct 60gccggcttag gacccccagc tccgacatgt cgccctctgg tcgcctgtgt cttctcacca 120tcgttggcct gattctcccc accagaggac agacgttgaa agataccacg tccagttctt 180cagcagactc aactatcatg gacattcagg tcccgacacg agccccagat gcagtctaca 240cagaactcca gcccacctct ccaaccccaa cctggcctgc tgatgaaaca ccacaacccc 300agacccagac ccagcaactg gaaggaacgg atgggcctct agtgacagat ccagagacac 360acaagagcac caaagcagct catcccactg atgacaccac gacgctctct gagagaccat 420ccccaagcac agacgtccag acagaccccc agaccctcaa gccatctggt tttcatgagg 480atgacccctt cttctatgat gaacacaccc tccggaaacg ggggctgttg gtcgcagctg 540tgctgttcat cacaggcatc atcatcctca ccagtggcaa gtgcaggcag ctgtcccggt 600tatgccggaa tcgttgcagg tgagtccatc agaaacagga gctgacaacc tgctgggcac 660ccgaagacca agccccctgc cagctcaccg tgcccagcct cctgcatccc ctcgaagagc 720ctggccagag agggaagaca cagatgatga agctggagcc agggctgccg gtccgagtct 780cctacctccc ccaaccctgc ccgcccctga aggctacctg gcgccttggg ggctgtccct 840caagttatct cctctgctaa gacaaaaagt aaagcactgt ggtctttgcc 890211104DNAHomo sapiens 21gcgcgagtga aggaagacga agtgcgtgac ccgaccggct gtggtgttcc agtccccact 60gaccagtagg agcagcaggg cgtcggcttg tgaggtggct tttcctcggg gcaacccagg 120aaggccccaa gaggacaatg gattctggaa ctcgcccagt tggtagctgc tgtagcagcc 180ccgctgggct ctcacgggag tacaaactag tgatgctggg tgctggtggt gtagggaaga 240gtgccatgac catgcagttc atcagccacc gattcccaga agatcatgat cccaccattg 300aagatgctta taagatcagg atccgtattg atgatgagcc tgccaatctg gacattttgg 360atacagctgg acaggcagag tttacagcca tgcgggacca gtatatgagg gcaggagaag 420ggtttatcat ctgttactct atcacggatc gtcgaagttt ccatgaagtt cgwgagttta 480aacagcttat ttatcgagtc cgacgtactg acgatacacc tgtggttctt gtgggaaaca 540agtcagacct caaacagcta agacaggtca ccaaggaaga aggattggcc ttggcccgag 600aattcagctg tccctttttt gagacatctg ctgcataccg ctactatatt gatgatgttt 660tccatgccct tgtacgggag atacgtagga aagaaaagga ggcagtactg gccatggaga 720aaaaatctaa gcccaaaaac agtgtatgga agaggctaaa atcaccattc cggaagaaga 780aagattcagt aacttgaaga gaagatgtga agtgtttatc tgtgaactgc agtgctgtat 840caaagcagtc cagtaacctg cagtactgag tatggtgctt gctctttcac ttaactgata 900agagggacat gcctactagg agtttttaat gatgtggtat ttaaagtatt gtctcttagt 960taagtatgat ttattaaccc agtggagcac tgtctgcttt taaattgtca cattagaatt 1020tgttctacca atgttttggg ttctgttgcg ctattaatta atgtaaattt gtttataccc 1080aggagaaaaa aaaaaaaaaa aaaa 1104223651DNAHomo sapiens 22tgaagattca gttttcactt aaacaaccag caagtcttga agtctcttcc caagcaaatg 60ggagcttctt tggaccttgg agcacacaga ggattctact ttctttaaaa ctttgttttc 120aggcaatttc cctgagaacc gtttacttcc agaagattgg tggagcttga tctgaaggct 180ggccatgaaa tctcaaggtc aacattggta ttccagttca gataaaaact gtaaagtgag 240ctttcgtgag aagcttctga ttattgattc aaacctgggg gtccaagatg tggagaacct 300caagtttctc tgcataggat tggtccccaa caagaagctg gagaagtcca gctcagcctc 360agatgttttt gaacatctct tggcagagga tctgctgagt gaggaagacc ctttcttcct 420ggcagaactc ctctatatca tacggcagaa gaagctgctg cagcacctca actgtaccaa 480agaggaagtg gagcgactgc tgcccacccg acaaagggtt tctctgttta gaaacctgct 540ctacgaactg tcagaaggca ttgactcaga gaacttaaag gacatgatct tccttctgaa 600agactcgctt cccaaaactg aaatgacctc cctaagtttc ctggcatttc tagagaaaca 660aggtaaaata gatgaagata atctgacatg cctggaggac ctctgcaaaa cagttgtacc 720taaacttttg agaaacatag agaaatacaa aagagagaaa gctatccaga tagtgacacc 780tcctgtagac aaggaagccg agtcgtatca aggagaggaa gaactagttt cccaaacaga 840tgttaagaca ttcttggaag ccttaccgag ggcagctgtg tacaggatga atcggaacca 900cagaggcctc tgtgtcattg tcaacaacca cagctttacc tccctgaagg acagacaagg 960aacccataaa gatgctgaga tcctgagtca tgtgttccag tggcttgggt tcacagtgca 1020tatacacaat aatgtgacga aagtggaaat ggagatggtc ctgcagaagc agaagtgcaa 1080tccagcccat gccgacgggg actgcttcgt gttctgtatt ctgacccatg ggagatttgg 1140agctgtctac tcttcggatg aggccctcat tcccattcgg gagatcatgt ctcacttcac 1200agccctgcag tgccctagac tggctgaaaa acctaaactc tttttcatcc aggcctgcca 1260aggtgaagag atacagcctt ccgtatccat cgaagcagat gctctgaacc ctgagcaggc 1320acccacttcc ctgcaggaca gtattcctgc cgaggctgac ttcctacttg gtctggccac 1380tgtcccaggc tatgtatcct ttcggcatgt ggaggaaggc agctggtata ttcagtctct 1440gtgtaatcat ctgaagaaat tggtcccaag acatgaagac atcttatcca tcctcactgc 1500tgtcaacgat gatgtgagtc gaagagtgga caaacaggga acaaagaaac agatgcccca 1560gcctgctttc acactaagga aaaaactagt attccctgtg cccctggatg cactttcatt 1620atagcagaga gtttttgttg gttcttagac ctcaaacgaa tcattggcta taacctccag 1680cctcctgccc agcacaggaa tcggtggtct ccacctgtca ttctagaaac aggaaacacc 1740gtgttttctg acacagtcaa ttctgatttt ctttttcttt tgcaagtcta aatgttagaa 1800aactttcttt tttttggaga tagtctcatt ctgtcaccca gactggagtg caggggggca 1860atcacggctc actgtagtct cgacctccca ggctcaagct gtcctcccac ctcagcctcc 1920caagtagctg agactacagg tgtgtgtcca tgcacagcta actttttatt ttttttgtgg 1980agatggggtt tcactatgtt gcctaagctg gtctcaaact cctgggctca agcgatcctc 2040ccacctcagc ttctcaaagt tctgggacta caggcatgaa atactgtgcc tggcctgggg 2100accaggtgca ttttaaggtt ccttggtgtt caaaaaccac gttcttagcc tagattgagc 2160ttagattgcc tctctagaca actacccctt agttataatt ctgtgtcccc tctgcatgcc 2220cttaaacatt ggacagtgag gtcacagtcc acccaccctc tctctgatct cccccttcct 2280aagacttctc ttttgcacat ctagtgaggt gaaaatttgg tctatgccag gcccatttcc 2340tgcttttgtg taaggaaggt gctcacatag gaagttttta tttggttaga gacaggtttc 2400cctgtaggaa gatgatggct catttacact cagctgctct gcaagcagaa actttacaac 2460ctgatgtcat attccatttt ggactgggtg cggtgactca tgcctgtaat cccagtactc 2520tgggaagcca aggcaggcag atcacttgag gtcaggagtt cgagaccagc ctggccaata 2580cggcaaaacc tcatcattac taaaaacaca aaaattagcc aggtgtggcg gcgagcacct 2640gtaatcccag ctactcggga ggctgagaca ggagaatctc ttgaatccag gaggcagagg 2700ctgtggtgag ccaagatgac acaactgcac tccagcttgg gcaacagggc gagaccttgt 2760ttaaaaaaaa aattcaatat tggggttgga acatttcagt tgccattgac agaacaccca 2820attcaaattg actgaagcaa agaagggaat ttattgcctc tttcacattg aaacccagga 2880gtggataaca ctggcttcag gcaaagcttg aatcaggact caatctacag gccagcacct 2940ttctcttggc cggatgtcct cagggctggc agatgcagta gactgcagtg gacagtcccc 3000accttgttac tgctactaca ctttgctcct ctggcccaag gcatgaggag agaggctgtg 3060tcagaaactg aagctgttct caggatcact gggctcttct tggcagaggg gatgtctggc 3120ttgcctgaag ggagtggctc tgtaaggacg ccttgatgct ttcttcatta agattttgag 3180catttttacg tacttgagct tttttttttt tttttttcaa tttctagagg aactttttct 3240ctgttaattc ctggaactgt attttgaatc cttaaaggtg agccctcata gggagatcca 3300aagtcctgtg gttaacgcct tcatttatag atgaggcagc tgaggcctgg ggatgtgaac 3360aacctgctca cagtcctcat ttactggatt tgacttcagc caggtgaact ggaatgcctt 3420ggggcgtgga agggcattag gagtgtttca tttgatatgt gaatgctcat aaaaaaatgt 3480caaggaatga agaacaacaa ctctcagtgg tgcctgcatt tataattatt tatgtgaaag 3540tcaaattcat gtacagtaaa tttgttataa gaatattcac aagaacactg ttctgatatc 3600tctgattgtc atgtggattt gaatgtagct tgacagagaa aaaaaaaaaa a 3651232054DNAHomo sapiens 23tgaagattca gttttcactt aaacaaccag caagtcttga agtctcttcc caagcaaatg 60ggagcttctt tggaccttgg agcacacaga ggattctact ttctttaaaa ctttgttttc 120aggcaatttc cctgagaacc gtttacttcc agaagattgg tggagcttga tctgaaggct 180ggccatgaaa tctcaaggtc aacattggta ttccagttca gataaaaact gtaaagtgag 240ctttcgtgag aagcttctga ttattgattc aaacctgggg gtccaagatg tggagaacct 300caagtttctc tgcataggat tggtccccaa caagaagctg gagaagtcca gctcagcctc 360agatgttttt gaacatctct tggcagagga tctgctgagt gaggaagacc ctttcttcct 420ggcagaactc ctctatatca tacggcagaa gaagctgctg cagcacctca actgtaccaa 480agaggaagtg gagcgactgc tgcccacccg acaaagggtt tctctgttta gaaacctgct 540ctacgaactg tcagaaggca ttgactcaga gaacttaaag gacatgatct tccttctgaa 600agactcgctt cccaaaactg aaatgacctc cctaagtttc ctggcatttc tagagaaaca 660aggtaaaata gatgaagata atctgacatg cctggaggac ctctgcaaaa cagttgtacc 720taaacttttg agaaacatag agaaatacaa aagagagaaa gctatccaga tagtgacacc 780tcctgtagac aaggaagccg agtcgtatca aggagaggaa gaactagttt cccaaacaga 840tgttaagaca ttcttggaag ccttaccgca ggagtcctgg caaaataagc atgcaggtag 900taatggtaac agagccacaa atggtgcacc aagcctggtc tccaggggga tgcaaggagc 960atctgctaac actctaaact ctgaaaccag cacaaagagg gcagctgtgt acaggatgaa 1020tcggaaccac agaggcctct gtgtcattgt caacaaccac agctttacct ccctgaagga 1080cagacaagga acccataaag atgctgagat cctgagtcat gtgttccagt ggcttgggtt 1140cacagtgcat atacacaata atgtgacgaa agtggaaatg gagatggtcc tgcagaagca 1200gaagtgcaat ccagcccatg ccgacgggga ctgcttcgtg ttctgtattc tgacccatgg 1260gagatttgga gctgtctact cttcggatga ggccctcatt cccattcggg agatcatgtc 1320tcacttcaca gccctgcagt gccctagact ggctgaaaaa cctaaactct ttttcatcca 1380ggcctgccaa ggtgaagaga tacagccttc cgtatccatc gaagcagatg ctctgaaccc 1440tgagcaggca cccacttccc tgcaggacag tattcctgcc gaggctgact tcctacttgg 1500tctggccact gtcccaggct atgtatcctt tcggcatgtg gaggaaggca gctggtatat 1560tcagtctctg tgtaatcatc tgaagaaatt ggtcccaagg atgctgaaat ttctggaaaa 1620gacaatggaa atcaggggca ggaagagaac agtgtggggt gctaaacaga tctcagcaac 1680ctccctgccc acggccatct ctgcgcagac acctcgaccc cccatgcgca ggtggagcag 1740cgtttcctag ttctttccag aggcttcctt ctgcctgcct tccagccaca tcgcctgaga 1800ttgacaacgc cctacagcaa gacggaaacc tccctttaca gcaccacctt gcgattctgc 1860agccacaaag ttgagacttc tgaacgtggc actcttctgt tcccttactg tttcacgtgt 1920acctgtgtca tctttcttgt ttcatcgtaa acatacttct aaaattccca ttttctttat 1980ttagaaatag aactacaagc ggatggttaa acaatttaaa caaatggtcc atggggaaaa 2040gtgaatttca cact 2054243688DNAHomo sapiens 24tgaagattca gttttcactt aaacaaccag caagtcttga agtctcttcc caagcaaatg 60ggagcttctt tggaccttgg agcacacaga ggattctact ttctttaaaa ctttgttttc 120aggcaatttc cctgagaacc gtttacttcc agaagattgg tggagcttga tctgaaggct 180ggccatgaaa tctcaaggtc aacattggta ttccagttca gataaaaact gtaaagtgag 240ctttcgtgag aagcttctga ttattgattc aaacctgggg gtccaagatg tggagaacct 300caagtttctc tgcataggat tggtccccaa caagaagctg gagaagtcca gctcagcctc 360agatgttttt gaacatctct tggcagagga tctgctgagt gaggaagacc ctttcttcct 420ggcagaactc ctctatatca tacggcagaa gaagctgctg cagcacctca actgtaccaa 480agaggaagtg gagcgactgc tgcccacccg acaaagggtt tctctgttta gaaacctgct 540ctacgaactg tcagaaggca ttgactcaga gaacttaaag gacatgatct tccttctgaa 600agactcgctt cccaaaactg aaatgacctc cctaagtttc ctggcatttc tagagaaaca 660aggtaaaata gatgaagata atctgacatg cctggaggac ctctgcaaaa cagttgtacc 720taaacttttg agaaacatag agaaatacaa aagagagaaa gctatccaga tagtgacacc 780tcctgtagac aaggaagccg agtcgtatca aggagaggaa gaactagttt cccaaacaga 840tgttaagaca ttcttggaag ccttaccgca ggagtcctgg caaaataagc atgcaggtag 900taatgagggc agctgtgtac aggatgaatc ggaaccacag aggcctctgt gtcattgtca 960acaaccacag ctttacctcc ctgaaggaca gacaaggaac ccataaagat gctgagatcc 1020tgagtcatgt gttccagtgg cttgggttca cagtgcatat acacaataat gtgacgaaag 1080tggaaatgga gatggtcctg cagaagcaga agtgcaatcc agcccatgcc gacggggact 1140gcttcgtgtt ctgtattctg acccatggga gatttggagc tgtctactct tcggatgagg 1200ccctcattcc cattcgggag atcatgtctc acttcacagc cctgcagtgc cctagactgg 1260ctgaaaaacc taaactcttt ttcatccagg cctgccaagg tgaagagata cagccttccg 1320tatccatcga agcagatgct ctgaaccctg agcaggcacc cacttccctg caggacagta 1380ttcctgccga ggctgacttc ctacttggtc tggccactgt cccaggctat gtatcctttc 1440ggcatgtgga ggaaggcagc tggtatattc agtctctgtg taatcatctg aagaaattgg 1500tcccaagaca tgaagacatc ttatccatcc tcactgctgt caacgatgat gtgagtcgaa 1560gagtggacaa acagggaaca aagaaacaga tgccccagcc tgctttcaca ctaaggaaaa 1620aactagtatt ccctgtgccc ctggatgcac tttcattata gcagagagtt tttgttggtt 1680cttagacctc aaacgaatca ttggctataa cctccagcct cctgcccagc acaggaatcg 1740gtggtctcca cctgtcattc tagaaacagg aaacaccgtg ttttctgaca cagtcaattc 1800tgattttctt tttcttttgc aagtctaaat gttagaaaac tttctttttt ttggagatag 1860tctcattctg tcacccagac tggagtgcag gggggcaatc acggctcact gtagtctcga 1920cctcccaggc tcaagctgtc ctcccacctc agcctcccaa gtagctgaga ctacaggtgt 1980gtgtccatgc acagctaact ttttattttt tttgtggaga tggggtttca ctatgttgcc 2040taagctggtc tcaaactcct gggctcaagc gatcctccca cctcagcttc tcaaagttct 2100gggactacag gcatgaaata ctgtgcctgg cctggggacc aggtgcattt taaggttcct 2160tggtgttcaa aaaccacgtt cttagcctag attgagctta gattgcctct ctagacaact 2220accccttagt tataattctg tgtcccctct gcatgccctt aaacattgga cagtgaggtc 2280acagtccacc caccctctct ctgatctccc ccttcctaag acttctcttt tgcacatcta 2340gtgaggtgaa aatttggtct atgccaggcc catttcctgc ttttgtgtaa ggaaggtgct 2400cacataggaa gtttttattt ggttagagac aggtttccct gtaggaagat gatggctcat 2460ttacactcag ctgctctgca agcagaaact ttacaacctg atgtcatatt ccattttgga 2520ctgggtgcgg tgactcatgc ctgtaatccc agtactctgg gaagccaagg caggcagatc 2580acttgaggtc aggagttcga gaccagcctg gccaatacgg caaaacctca tcattactaa 2640aaacacaaaa attagccagg tgtggcggcg agcacctgta atcccagcta ctcgggaggc 2700tgagacagga gaatctcttg aatccaggag gcagaggctg tggtgagcca agatgacaca 2760actgcactcc agcttgggca acagggcgag accttgttta aaaaaaaaat tcaatattgg 2820ggttggaaca tttcagttgc cattgacaga acacccaatt caaattgact gaagcaaaga 2880agggaattta ttgcctcttt cacattgaaa cccaggagtg gataacactg gcttcaggca 2940aagcttgaat caggactcaa tctacaggcc agcacctttc tcttggccgg atgtcctcag 3000ggctggcaga tgcagtagac tgcagtggac agtccccacc ttgttactgc tactacactt 3060tgctcctctg gcccaaggca tgaggagaga ggctgtgtca gaaactgaag ctgttctcag 3120gatcactggg ctcttcttgg cagaggggat gtctggcttg cctgaaggga gtggctctgt 3180aaggacgcct tgatgctttc ttcattaaga ttttgagcat ttttacgtac ttgagctttt 3240tttttttttt ttttcaattt ctagaggaac tttttctctg ttaattcctg gaactgtatt 3300ttgaatcctt aaaggtgagc cctcataggg agatccaaag tcctgtggtt aacgccttca 3360tttatagatg aggcagctga ggcctgggga tgtgaacaac ctgctcacag tcctcattta 3420ctggatttga cttcagccag gtgaactgga atgccttggg gcgtggaagg gcattaggag 3480tgtttcattt gatatgtgaa tgctcataaa aaaatgtcaa ggaatgaaga acaacaactc 3540tcagtggtgc ctgcatttat aattatttat gtgaaagtca aattcatgta cagtaaattt 3600gttataagaa tattcacaag aacactgttc tgatatctct gattgtcatg tggatttgaa 3660tgtagcttga cagagaaaaa aaaaaaaa 3688253780DNAHomo sapiens 25tgaagattca gttttcactt aaacaaccag caagtcttga agtctcttcc caagcaaatg 60ggagcttctt tggaccttgg agcacacaga ggattctact ttctttaaaa ctttgttttc 120aggcaatttc cctgagaacc gtttacttcc agaagattgg tggagcttga tctgaaggct 180ggccatgaaa tctcaaggtc aacattggta ttccagttca gataaaaact gtaaagtgag 240ctttcgtgag aagcttctga ttattgattc aaacctgggg gtccaagatg tggagaacct 300caagtttctc tgcataggat tggtccccaa caagaagctg gagaagtcca gctcagcctc 360agatgttttt gaacatctct tggcagagga tctgctgagt gaggaagacc ctttcttcct 420ggcagaactc ctctatatca tacggcagaa gaagctgctg cagcacctca actgtaccaa 480agaggaagtg gagcgactgc tgcccacccg acaaagggtt tctctgttta gaaacctgct 540ctacgaactg tcagaaggca ttgactcaga gaacttaaag gacatgatct tccttctgaa 600agactcgctt cccaaaactg aaatgacctc cctaagtttc ctggcatttc tagagaaaca 660aggtaaaata gatgaagata atctgacatg cctggaggac ctctgcaaaa cagttgtacc 720taaacttttg agaaacatag agaaatacaa aagagagaaa gctatccaga tagtgacacc 780tcctgtagac aaggaagccg agtcgtatca aggagaggaa gaactagttt cccaaacaga 840tgttaagaca ttcttggaag ccttaccgca ggagtcctgg caaaataagc atgcaggtag 900taatggtaac agagccacaa atggtgcacc aagcctggtc tccaggggga tgcaaggagc 960atctgctaac actctaaact ctgaaaccag cacaaagagg gcagctgtgt acaggatgaa 1020tcggaaccac agaggcctct gtgtcattgt caacaaccac agctttacct ccctgaagga 1080cagacaagga acccataaag atgctgagat cctgagtcat gtgttccagt ggcttgggtt 1140cacagtgcat atacacaata atgtgacgaa agtggaaatg gagatggtcc tgcagaagca 1200gaagtgcaat ccagcccatg ccgacgggga ctgcttcgtg ttctgtattc tgacccatgg 1260gagatttgga gctgtctact cttcggatga ggccctcatt cccattcggg agatcatgtc 1320tcacttcaca gccctgcagt gccctagact ggctgaaaaa cctaaactct ttttcatcca 1380ggcctgccaa ggtgaagaga tacagccttc cgtatccatc gaagcagatg ctctgaaccc 1440tgagcaggca cccacttccc tgcaggacag tattcctgcc gaggctgact tcctacttgg 1500tctggccact gtcccaggct atgtatcctt tcggcatgtg gaggaaggca gctggtatat 1560tcagtctctg tgtaatcatc tgaagaaatt ggtcccaaga catgaagaca tcttatccat 1620cctcactgct gtcaacgatg atgtgagtcg aagagtggac aaacagggaa caaagaaaca 1680gatgccccag cctgctttca cactaaggaa aaaactagta ttccctgtgc ccctggatgc 1740actttcatta tagcagagag tttttgttgg ttcttagacc
tcaaacgaat cattggctat 1800aacctccagc ctcctgccca gcacaggaat cggtggtctc cacctgtcat tctagaaaca 1860ggaaacaccg tgttttctga cacagtcaat tctgattttc tttttctttt gcaagtctaa 1920atgttagaaa actttctttt ttttggagat agtctcattc tgtcacccag actggagtgc 1980aggggggcaa tcacggctca ctgtagtctc gacctcccag gctcaagctg tcctcccacc 2040tcagcctccc aagtagctga gactacaggt gtgtgtccat gcacagctaa ctttttattt 2100tttttgtgga gatggggttt cactatgttg cctaagctgg tctcaaactc ctgggctcaa 2160gcgatcctcc cacctcagct tctcaaagtt ctgggactac aggcatgaaa tactgtgcct 2220ggcctgggga ccaggtgcat tttaaggttc cttggtgttc aaaaaccacg ttcttagcct 2280agattgagct tagattgcct ctctagacaa ctacccctta gttataattc tgtgtcccct 2340ctgcatgccc ttaaacattg gacagtgagg tcacagtcca cccaccctct ctctgatctc 2400ccccttccta agacttctct tttgcacatc tagtgaggtg aaaatttggt ctatgccagg 2460cccatttcct gcttttgtgt aaggaaggtg ctcacatagg aagtttttat ttggttagag 2520acaggtttcc ctgtaggaag atgatggctc atttacactc agctgctctg caagcagaaa 2580ctttacaacc tgatgtcata ttccattttg gactgggtgc ggtgactcat gcctgtaatc 2640ccagtactct gggaagccaa ggcaggcaga tcacttgagg tcaggagttc gagaccagcc 2700tggccaatac ggcaaaacct catcattact aaaaacacaa aaattagcca ggtgtggcgg 2760cgagcacctg taatcccagc tactcgggag gctgagacag gagaatctct tgaatccagg 2820aggcagaggc tgtggtgagc caagatgaca caactgcact ccagcttggg caacagggcg 2880agaccttgtt taaaaaaaaa attcaatatt ggggttggaa catttcagtt gccattgaca 2940gaacacccaa ttcaaattga ctgaagcaaa gaagggaatt tattgcctct ttcacattga 3000aacccaggag tggataacac tggcttcagg caaagcttga atcaggactc aatctacagg 3060ccagcacctt tctcttggcc ggatgtcctc agggctggca gatgcagtag actgcagtgg 3120acagtcccca ccttgttact gctactacac tttgctcctc tggcccaagg catgaggaga 3180gaggctgtgt cagaaactga agctgttctc aggatcactg ggctcttctt ggcagagggg 3240atgtctggct tgcctgaagg gagtggctct gtaaggacgc cttgatgctt tcttcattaa 3300gattttgagc atttttacgt acttgagctt tttttttttt ttttttcaat ttctagagga 3360actttttctc tgttaattcc tggaactgta ttttgaatcc ttaaaggtga gccctcatag 3420ggagatccaa agtcctgtgg ttaacgcctt catttataga tgaggcagct gaggcctggg 3480gatgtgaaca acctgctcac agtcctcatt tactggattt gacttcagcc aggtgaactg 3540gaatgccttg gggcgtggaa gggcattagg agtgtttcat ttgatatgtg aatgctcata 3600aaaaaatgtc aaggaatgaa gaacaacaac tctcagtggt gcctgcattt ataattattt 3660atgtgaaagt caaattcatg tacagtaaat ttgttataag aatattcaca agaacactgt 3720tctgatatct ctgattgtca tgtggatttg aatgtagctt gacagagaaa aaaaaaaaaa 3780268RNAArtificial SequenceLinker sequence 26uugcuaua 827540PRTHomo sapiens 27Met Asn Gly Glu Ala Ile Cys Ser Ala Leu Pro Thr Ile Pro Tyr His1 5 10 15Lys Leu Ala Asp Leu Arg Tyr Leu Ser Arg Gly Ala Ser Gly Thr Val 20 25 30Ser Ser Ala Arg His Ala Asp Trp Arg Val Gln Val Ala Val Lys His 35 40 45Leu His Ile His Thr Pro Leu Leu Asp Ser Glu Arg Lys Asp Val Leu 50 55 60Arg Glu Ala Glu Ile Leu His Lys Ala Arg Phe Ser Tyr Ile Leu Pro65 70 75 80Ile Leu Gly Ile Cys Asn Glu Pro Glu Phe Leu Gly Ile Val Thr Glu 85 90 95Tyr Met Pro Asn Gly Ser Leu Asn Glu Leu Leu His Arg Lys Thr Glu 100 105 110Tyr Pro Asp Val Ala Trp Pro Leu Arg Phe Arg Ile Leu His Glu Ile 115 120 125Ala Leu Gly Val Asn Tyr Leu His Asn Met Thr Pro Pro Leu Leu His 130 135 140His Asp Leu Lys Thr Gln Asn Ile Leu Leu Asp Asn Glu Phe His Val145 150 155 160Lys Ile Ala Asp Phe Gly Leu Ser Lys Trp Arg Met Met Ser Leu Ser 165 170 175Gln Ser Arg Ser Ser Lys Ser Ala Pro Glu Gly Gly Thr Ile Ile Tyr 180 185 190Met Pro Pro Glu Asn Tyr Glu Pro Gly Gln Lys Ser Arg Ala Ser Ile 195 200 205Lys His Asp Ile Tyr Ser Tyr Ala Val Ile Thr Trp Glu Val Leu Ser 210 215 220Arg Lys Gln Pro Phe Glu Asp Val Thr Asn Pro Leu Gln Ile Met Tyr225 230 235 240Ser Val Ser Gln Gly His Arg Pro Val Ile Asn Glu Glu Ser Leu Pro 245 250 255Tyr Asp Ile Pro His Arg Ala Arg Met Ile Ser Leu Ile Glu Ser Gly 260 265 270Trp Ala Gln Asn Pro Asp Glu Arg Pro Ser Phe Leu Lys Cys Leu Ile 275 280 285Glu Leu Glu Pro Val Leu Arg Thr Phe Glu Glu Ile Thr Phe Leu Glu 290 295 300Ala Val Ile Gln Leu Lys Lys Thr Lys Leu Gln Ser Val Ser Ser Ala305 310 315 320Ile His Leu Cys Asp Lys Lys Lys Met Glu Leu Ser Leu Asn Ile Pro 325 330 335Val Asn His Gly Pro Gln Glu Glu Ser Cys Gly Ser Ser Gln Leu His 340 345 350Glu Asn Ser Gly Ser Pro Glu Thr Ser Arg Ser Leu Pro Ala Pro Gln 355 360 365Asp Asn Asp Phe Leu Ser Arg Lys Ala Gln Asp Cys Tyr Phe Met Lys 370 375 380Leu His His Cys Pro Gly Asn His Ser Trp Asp Ser Thr Ile Ser Gly385 390 395 400Ser Gln Arg Ala Ala Phe Cys Asp His Lys Thr Thr Pro Cys Ser Ser 405 410 415Ala Ile Ile Asn Pro Leu Ser Thr Ala Gly Asn Ser Glu Arg Leu Gln 420 425 430Pro Gly Ile Ala Gln Gln Trp Ile Gln Ser Lys Arg Glu Asp Ile Val 435 440 445Asn Gln Met Thr Glu Ala Cys Leu Asn Gln Ser Leu Asp Ala Leu Leu 450 455 460Ser Arg Asp Leu Ile Met Lys Glu Asp Tyr Glu Leu Val Ser Thr Lys465 470 475 480Pro Thr Arg Thr Ser Lys Val Arg Gln Leu Leu Asp Thr Thr Asp Ile 485 490 495Gln Gly Glu Glu Phe Ala Lys Val Ile Val Gln Lys Leu Lys Asp Asn 500 505 510Lys Gln Met Gly Leu Gln Pro Tyr Pro Glu Ile Leu Val Val Ser Arg 515 520 525Ser Pro Ser Leu Asn Leu Leu Gln Asn Lys Ser Met 530 535 54028737PRTHomo sapiens 28Met Val Val Phe Asn Gly Leu Leu Lys Ile Lys Ile Cys Glu Ala Val1 5 10 15Ser Leu Lys Pro Thr Ala Trp Ser Leu Arg His Ala Val Gly Pro Arg 20 25 30Pro Gln Thr Phe Leu Leu Asp Pro Tyr Ile Ala Leu Asn Val Asp Asp 35 40 45Ser Arg Ile Gly Gln Thr Ala Thr Lys Gln Lys Thr Asn Ser Pro Ala 50 55 60Trp His Asp Glu Phe Val Thr Asp Val Cys Asn Gly Arg Lys Ile Glu65 70 75 80Leu Ala Val Phe His Asp Ala Pro Ile Gly Tyr Asp Asp Phe Val Ala 85 90 95Asn Cys Thr Ile Gln Phe Glu Glu Leu Leu Gln Asn Gly Ser Arg His 100 105 110Phe Glu Asp Trp Ile Asp Leu Glu Pro Glu Gly Arg Val Tyr Val Ile 115 120 125Ile Asp Leu Ser Gly Ser Ser Gly Glu Ala Pro Lys Asp Asn Glu Glu 130 135 140Arg Val Phe Arg Glu Arg Met Arg Pro Arg Lys Arg Gln Gly Ala Val145 150 155 160Arg Arg Arg Val His Gln Val Asn Gly His Lys Phe Met Ala Thr Tyr 165 170 175Leu Arg Gln Pro Thr Tyr Cys Ser His Cys Arg Asp Phe Ile Trp Gly 180 185 190Val Ile Gly Lys Gln Gly Tyr Gln Cys Gln Val Cys Thr Cys Val Val 195 200 205His Lys Arg Cys His Glu Leu Ile Ile Thr Lys Cys Ala Gly Leu Lys 210 215 220Lys Gln Glu Thr Pro Asp Gln Val Gly Ser Gln Arg Phe Ser Val Asn225 230 235 240Met Pro His Lys Phe Gly Ile His Asn Tyr Lys Val Pro Thr Phe Cys 245 250 255Asp His Cys Gly Ser Leu Leu Trp Gly Leu Leu Arg Gln Gly Leu Gln 260 265 270Cys Lys Val Cys Lys Met Asn Val His Arg Arg Cys Glu Thr Asn Val 275 280 285Ala Pro Asn Cys Gly Val Asp Ala Arg Gly Ile Ala Lys Val Leu Ala 290 295 300Asp Leu Gly Val Thr Pro Asp Lys Ile Thr Asn Ser Gly Gln Arg Arg305 310 315 320Lys Lys Leu Ile Ala Gly Ala Glu Ser Pro Gln Pro Ala Ser Gly Ser 325 330 335Ser Pro Ser Glu Glu Asp Arg Ser Lys Ser Ala Pro Thr Ser Pro Cys 340 345 350Asp Gln Glu Ile Lys Glu Leu Glu Asn Asn Ile Arg Lys Ala Leu Ser 355 360 365Phe Asp Asn Arg Gly Glu Glu His Arg Ala Ala Ser Ser Pro Asp Gly 370 375 380Gln Leu Met Ser Pro Gly Glu Asn Gly Glu Val Arg Gln Gly Gln Ala385 390 395 400Lys Arg Leu Gly Leu Asp Glu Phe Asn Phe Ile Lys Val Leu Gly Lys 405 410 415Gly Ser Phe Gly Lys Val Met Leu Ala Glu Leu Lys Gly Lys Asp Glu 420 425 430Val Tyr Ala Val Lys Val Leu Lys Lys Asp Val Ile Leu Gln Asp Asp 435 440 445Asp Val Asp Cys Thr Met Thr Glu Lys Arg Ile Leu Ala Leu Ala Arg 450 455 460Lys His Pro Tyr Leu Thr Gln Leu Tyr Cys Cys Phe Gln Thr Lys Asp465 470 475 480Arg Leu Phe Phe Val Met Glu Tyr Val Asn Gly Gly Asp Leu Met Phe 485 490 495Gln Ile Gln Arg Ser Arg Lys Phe Asp Glu Pro Arg Ser Arg Phe Tyr 500 505 510Ala Ala Glu Val Thr Ser Ala Leu Met Phe Leu His Gln His Gly Val 515 520 525Ile Tyr Arg Asp Leu Lys Leu Asp Asn Ile Leu Leu Asp Ala Glu Gly 530 535 540His Cys Lys Leu Ala Asp Phe Gly Met Cys Lys Glu Gly Ile Leu Asn545 550 555 560Gly Val Thr Thr Thr Thr Phe Cys Gly Thr Pro Asp Tyr Ile Ala Pro 565 570 575Glu Ile Leu Gln Glu Leu Glu Tyr Gly Pro Ser Val Asp Trp Trp Ala 580 585 590Leu Gly Val Leu Met Tyr Glu Met Met Ala Gly Gln Pro Pro Phe Glu 595 600 605Ala Asp Asn Glu Asp Asp Leu Phe Glu Ser Ile Leu His Asp Asp Val 610 615 620Leu Tyr Pro Val Trp Leu Ser Lys Glu Ala Val Ser Ile Leu Lys Ala625 630 635 640Phe Met Thr Lys Asn Pro His Lys Arg Leu Gly Cys Val Ala Ser Gln 645 650 655Asn Gly Glu Asp Ala Ile Lys Gln His Pro Phe Phe Lys Glu Ile Asp 660 665 670Trp Val Leu Leu Glu Gln Lys Lys Ile Lys Pro Pro Phe Lys Pro Arg 675 680 685Ile Lys Thr Lys Arg Asp Val Asn Asn Phe Asp Gln Asp Phe Thr Arg 690 695 700Glu Glu Pro Val Leu Thr Leu Val Asp Glu Ala Ile Val Lys Gln Ile705 710 715 720Asn Gln Glu Glu Phe Lys Gly Phe Ser Tyr Phe Gly Glu Asp Leu Met 725 730 735Pro29431PRTHomo sapiens 29Met Ala His Ser Pro Val Gln Ser Gly Leu Pro Gly Met Gln Asn Leu1 5 10 15Lys Ala Asp Pro Glu Glu Leu Phe Thr Lys Leu Glu Lys Ile Gly Lys 20 25 30Gly Ser Phe Gly Glu Val Phe Lys Gly Ile Asp Asn Arg Thr Gln Lys 35 40 45Val Val Ala Ile Lys Ile Ile Asp Leu Glu Glu Ala Glu Asp Glu Ile 50 55 60Glu Asp Ile Gln Gln Glu Ile Thr Val Leu Ser Gln Cys Asp Ser Pro65 70 75 80Tyr Val Thr Lys Tyr Tyr Gly Ser Tyr Leu Lys Asp Thr Lys Leu Trp 85 90 95Ile Ile Met Glu Tyr Leu Gly Gly Gly Ser Ala Leu Asp Leu Leu Glu 100 105 110Pro Gly Pro Leu Asp Glu Thr Gln Ile Ala Thr Ile Leu Arg Glu Ile 115 120 125Leu Lys Gly Leu Asp Tyr Leu His Ser Glu Lys Lys Ile His Arg Asp 130 135 140Ile Lys Ala Ala Asn Val Leu Leu Ser Glu His Gly Glu Val Lys Leu145 150 155 160Ala Asp Phe Gly Val Ala Gly Gln Leu Thr Asp Thr Gln Ile Lys Arg 165 170 175Asn Thr Phe Val Gly Thr Pro Phe Trp Met Ala Pro Glu Val Ile Lys 180 185 190Gln Ser Ala Tyr Asp Ser Lys Ala Asp Ile Trp Ser Leu Gly Ile Thr 195 200 205Ala Ile Glu Leu Ala Arg Gly Glu Pro Pro His Ser Glu Leu His Pro 210 215 220Met Lys Val Leu Phe Leu Ile Pro Lys Asn Asn Pro Pro Thr Leu Glu225 230 235 240Gly Asn Tyr Ser Lys Pro Leu Lys Glu Phe Val Glu Ala Cys Leu Asn 245 250 255Lys Glu Pro Ser Phe Arg Pro Thr Ala Lys Glu Leu Leu Lys His Lys 260 265 270Phe Ile Leu Arg Asn Ala Lys Lys Thr Ser Tyr Leu Thr Glu Leu Ile 275 280 285Asp Arg Tyr Lys Arg Trp Lys Ala Glu Gln Ser His Asp Asp Ser Ser 290 295 300Ser Glu Asp Ser Asp Ala Glu Thr Asp Gly Gln Ala Ser Gly Gly Ser305 310 315 320Asp Ser Gly Asp Trp Ile Phe Thr Ile Arg Glu Lys Asp Pro Lys Asn 325 330 335Leu Glu Asn Gly Ala Leu Gln Pro Ser Asp Leu Asp Arg Asn Lys Met 340 345 350Lys Asp Ile Pro Lys Arg Pro Phe Ser Gln Cys Leu Ser Thr Ile Ile 355 360 365Ser Pro Leu Phe Ala Glu Leu Lys Glu Lys Ser Gln Ala Cys Gly Gly 370 375 380Asn Leu Gly Ser Ile Glu Glu Leu Arg Gly Ala Ile Tyr Leu Ala Glu385 390 395 400Glu Val Cys Pro Gly Ile Ser Asp Thr Met Val Ala Gln Leu Val Gln 405 410 415Arg Leu Gln Arg Tyr Ser Leu Ser Gly Gly Gly Thr Ser Ser His 420 425 43030443PRTHomo sapiens 30Met Asp Ser Arg Ala Gln Leu Trp Gly Leu Ala Leu Asn Lys Arg Arg1 5 10 15Ala Thr Leu Pro His Pro Gly Gly Ser Thr Asn Leu Lys Ala Asp Pro 20 25 30Glu Glu Leu Phe Thr Lys Leu Glu Lys Ile Gly Lys Gly Ser Phe Gly 35 40 45Glu Val Phe Lys Gly Ile Asp Asn Arg Thr Gln Lys Val Val Ala Ile 50 55 60Lys Ile Ile Asp Leu Glu Glu Ala Glu Asp Glu Ile Glu Asp Ile Gln65 70 75 80Gln Glu Ile Thr Val Leu Ser Gln Cys Asp Ser Pro Tyr Val Thr Lys 85 90 95Tyr Tyr Gly Ser Tyr Leu Lys Asp Thr Lys Leu Trp Ile Ile Met Glu 100 105 110Tyr Leu Gly Gly Gly Ser Ala Leu Asp Leu Leu Glu Pro Gly Pro Leu 115 120 125Asp Glu Thr Gln Ile Ala Thr Ile Leu Arg Glu Ile Leu Lys Gly Leu 130 135 140Asp Tyr Leu His Ser Glu Lys Lys Ile His Arg Asp Ile Lys Ala Ala145 150 155 160Asn Val Leu Leu Ser Glu His Gly Glu Val Lys Leu Ala Asp Phe Gly 165 170 175Val Ala Gly Gln Leu Thr Asp Thr Gln Ile Lys Arg Asn Thr Phe Val 180 185 190Gly Thr Pro Phe Trp Met Ala Pro Glu Val Ile Lys Gln Ser Ala Tyr 195 200 205Asp Ser Lys Ala Asp Ile Trp Ser Leu Gly Ile Thr Ala Ile Glu Leu 210 215 220Ala Arg Gly Glu Pro Pro His Ser Glu Leu His Pro Met Lys Val Leu225 230 235 240Phe Leu Ile Pro Lys Asn Asn Pro Pro Thr Leu Glu Gly Asn Tyr Ser 245 250 255Lys Pro Leu Lys Glu Phe Val Glu Ala Cys Leu Asn Lys Glu Pro Ser 260 265 270Phe Arg Pro Thr Ala Lys Glu Leu Leu Lys His Lys Phe Ile Leu Arg 275 280 285Asn Ala Lys Lys Thr Ser Tyr Leu Thr Glu Leu Ile Asp Arg Tyr Lys 290 295 300Arg Trp Lys Ala Glu Gln Ser His Asp Asp Ser Ser Ser Glu Asp Ser305 310 315 320Asp Ala Glu Thr Asp Gly Gln Ala Ser Gly Gly Ser Asp Ser Gly Asp 325 330 335Trp Ile Phe Thr Ile Arg Glu Lys Asp Pro Lys Asn Leu Glu Asn Gly 340 345 350Ala Leu Gln Pro Ser Asp Leu Asp Arg Asn Lys Met Lys Asp Ile Pro 355 360 365Lys Arg Pro Phe Ser Gln Cys Leu Ser Thr Ile Ile Ser Pro Leu Phe 370 375 380Ala Glu Leu Lys Glu Lys Ser Gln Ala Cys Gly Gly Asn Leu Gly Ser385 390 395 400Ile Glu Glu Leu Arg Gly Ala Ile Tyr Leu Ala Glu Glu Ala Cys Pro
405 410 415Gly Ile Ser Asp Thr Met Val Ala Gln Leu Val Gln Arg Leu Gln Arg 420 425 430Tyr Ser Leu Ser Gly Gly Gly Thr Ser Ser His 435 44031471PRTHomo sapiens 31Met Ser Glu Glu Ser Asp Met Asp Lys Ala Ile Lys Glu Thr Ser Ile1 5 10 15Leu Glu Glu Tyr Ser Ile Asn Trp Thr Gln Lys Leu Gly Ala Gly Ile 20 25 30Ser Gly Pro Val Arg Val Cys Val Lys Lys Ser Thr Gln Glu Arg Phe 35 40 45Ala Leu Lys Ile Leu Leu Asp Arg Pro Lys Ala Arg Asn Glu Val Arg 50 55 60Leu His Met Met Cys Ala Thr His Pro Asn Ile Val Gln Ile Ile Glu65 70 75 80Val Phe Ala Asn Ser Val Gln Phe Pro His Glu Ser Ser Pro Arg Ala 85 90 95Arg Leu Leu Ile Val Met Glu Met Met Glu Gly Gly Glu Leu Phe His 100 105 110Arg Ile Ser Gln His Arg His Phe Thr Glu Lys Gln Ala Ser Gln Val 115 120 125Thr Lys Gln Ile Ala Leu Ala Leu Arg His Cys His Leu Leu Asn Ile 130 135 140Ala His Arg Asp Leu Lys Pro Glu Asn Leu Leu Phe Lys Asp Asn Ser145 150 155 160Leu Asp Ala Pro Val Lys Leu Cys Asp Phe Gly Phe Ala Lys Ile Asp 165 170 175Gln Gly Asp Leu Met Thr Pro Gln Phe Thr Pro Tyr Tyr Val Ala Pro 180 185 190Gln Val Leu Glu Ala Gln Arg Arg His Gln Lys Glu Lys Ser Gly Ile 195 200 205Ile Pro Thr Ser Pro Thr Pro Tyr Thr Tyr Asn Lys Ser Cys Asp Leu 210 215 220Trp Ser Leu Gly Val Ile Ile Tyr Val Met Leu Cys Gly Tyr Pro Pro225 230 235 240Phe Tyr Ser Lys His His Ser Arg Thr Ile Pro Lys Asp Met Arg Arg 245 250 255Lys Ile Met Thr Gly Ser Phe Glu Phe Pro Glu Glu Glu Trp Ser Gln 260 265 270Ile Ser Glu Met Ala Lys Asp Val Val Arg Lys Leu Leu Lys Val Lys 275 280 285Pro Glu Glu Arg Leu Thr Ile Glu Gly Val Leu Asp His Pro Trp Leu 290 295 300Asn Ser Thr Glu Ala Leu Asp Asn Val Leu Pro Ser Ala Gln Leu Met305 310 315 320Met Asp Lys Ala Val Val Ala Gly Ile Gln Gln Ala His Ala Glu Gln 325 330 335Leu Ala Asn Met Arg Ile Gln Asp Leu Lys Val Ser Leu Lys Pro Leu 340 345 350His Ser Val Asn Asn Pro Ile Leu Arg Lys Arg Lys Leu Leu Gly Thr 355 360 365Lys Pro Lys Asp Ser Val Tyr Ile His Asp His Glu Asn Gly Ala Glu 370 375 380Asp Ser Asn Val Ala Leu Glu Lys Leu Arg Asp Val Ile Ala Gln Cys385 390 395 400Ile Leu Pro Gln Ala Gly Glu Asn Glu Asp Glu Lys Leu Asn Glu Val 405 410 415Met Gln Glu Ala Trp Lys Tyr Asn Arg Glu Cys Lys Leu Leu Arg Asp 420 425 430Thr Leu Gln Ser Phe Ser Trp Asn Gly Arg Gly Phe Thr Asp Lys Val 435 440 445Asp Arg Leu Lys Leu Ala Glu Ile Val Lys Gln Val Ile Glu Glu Gln 450 455 460Thr Thr Ser His Glu Ser Gln465 47032473PRTHomo sapiens 32Met Ser Glu Glu Ser Asp Met Asp Lys Ala Ile Lys Glu Thr Ser Ile1 5 10 15Leu Glu Glu Tyr Ser Ile Asn Trp Thr Gln Lys Leu Gly Ala Gly Ile 20 25 30Ser Gly Pro Val Arg Val Cys Val Lys Lys Ser Thr Gln Glu Arg Phe 35 40 45Ala Leu Lys Ile Leu Leu Asp Arg Pro Lys Ala Arg Asn Glu Val Arg 50 55 60Leu His Met Met Cys Ala Thr His Pro Asn Ile Val Gln Ile Ile Glu65 70 75 80Val Phe Ala Asn Ser Val Gln Phe Pro His Glu Ser Ser Pro Arg Ala 85 90 95Arg Leu Leu Ile Val Met Glu Met Met Glu Gly Gly Glu Leu Phe His 100 105 110Arg Ile Ser Gln His Arg His Phe Thr Glu Lys Gln Ala Ser Gln Val 115 120 125Thr Lys Gln Ile Ala Leu Ala Leu Arg His Cys His Leu Leu Asn Ile 130 135 140Ala His Arg Asp Leu Lys Pro Glu Asn Leu Leu Phe Lys Asp Asn Ser145 150 155 160Leu Asp Ala Pro Val Lys Leu Cys Asp Phe Gly Phe Ala Lys Ile Asp 165 170 175Gln Gly Asp Leu Met Thr Pro Gln Phe Thr Pro Tyr Tyr Val Ala Pro 180 185 190Gln Val Leu Glu Ala Gln Arg Arg His Gln Lys Glu Lys Ser Gly Ile 195 200 205Ile Pro Thr Ser Pro Thr Pro Tyr Thr Tyr Asn Lys Ser Cys Asp Leu 210 215 220Trp Ser Leu Gly Val Ile Ile Tyr Val Met Leu Cys Gly Tyr Pro Pro225 230 235 240Phe Tyr Ser Lys His His Ser Arg Thr Ile Pro Lys Asp Met Arg Arg 245 250 255Lys Ile Met Thr Gly Ser Phe Glu Phe Pro Glu Glu Glu Trp Ser Gln 260 265 270Ile Ser Glu Met Ala Lys Asp Val Val Arg Lys Leu Leu Lys Val Lys 275 280 285Pro Glu Glu Arg Leu Thr Ile Glu Gly Val Leu Asp His Pro Trp Leu 290 295 300Asn Ser Thr Glu Ala Leu Asp Asn Val Leu Pro Ser Ala Gln Leu Met305 310 315 320Met Asp Lys Ala Val Val Ala Gly Ile Gln Gln Ala His Ala Glu Gln 325 330 335Leu Ala Asn Met Arg Ile Gln Asp Leu Lys Val Ser Leu Lys Pro Leu 340 345 350His Ser Val Asn Asn Pro Ile Leu Arg Lys Arg Lys Leu Leu Gly Thr 355 360 365Lys Pro Lys Asp Ser Val Tyr Ile His Asp His Glu Asn Gly Ala Glu 370 375 380Asp Ser Asn Val Ala Leu Glu Lys Leu Arg Asp Val Ile Ala Gln Cys385 390 395 400Ile Leu Pro Gln Ala Gly Lys Gly Glu Asn Glu Asp Glu Lys Leu Asn 405 410 415Glu Val Met Gln Glu Ala Trp Lys Tyr Asn Arg Glu Cys Lys Leu Leu 420 425 430Arg Asp Thr Leu Gln Ser Phe Ser Trp Asn Gly Arg Gly Phe Thr Asp 435 440 445Lys Val Asp Arg Leu Lys Leu Ala Glu Ile Val Lys Gln Val Ile Glu 450 455 460Glu Gln Thr Thr Ser His Glu Ser Gln465 47033465PRTHomo sapiens 33Met Val Ser Ser Gln Lys Leu Glu Lys Pro Ile Glu Met Gly Ser Ser1 5 10 15Glu Pro Leu Pro Ile Ala Asp Gly Asp Arg Arg Arg Lys Lys Lys Arg 20 25 30Arg Gly Arg Ala Thr Asp Ser Leu Pro Gly Lys Phe Glu Asp Met Tyr 35 40 45Lys Leu Thr Ser Glu Leu Leu Gly Glu Gly Ala Tyr Ala Lys Val Gln 50 55 60Gly Ala Val Ser Leu Gln Asn Gly Lys Glu Tyr Ala Val Lys Ile Ile65 70 75 80Glu Lys Gln Ala Gly His Ser Arg Ser Arg Val Phe Arg Glu Val Glu 85 90 95Thr Leu Tyr Gln Cys Gln Gly Asn Lys Asn Ile Leu Glu Leu Ile Glu 100 105 110Phe Phe Glu Asp Asp Thr Arg Phe Tyr Leu Val Phe Glu Lys Leu Gln 115 120 125Gly Gly Ser Ile Leu Ala His Ile Gln Lys Gln Lys His Phe Asn Glu 130 135 140Arg Glu Ala Ser Arg Val Val Arg Asp Val Ala Ala Ala Leu Asp Phe145 150 155 160Leu His Thr Lys Asp Lys Val Ser Leu Cys His Leu Gly Trp Ser Ala 165 170 175Met Ala Pro Ser Gly Leu Thr Ala Ala Pro Thr Ser Leu Gly Ser Ser 180 185 190Asp Pro Pro Thr Ser Ala Ser Gln Val Ala Gly Thr Thr Gly Ile Ala 195 200 205His Arg Asp Leu Lys Pro Glu Asn Ile Leu Cys Glu Ser Pro Glu Lys 210 215 220Val Ser Pro Val Lys Ile Cys Asp Phe Asp Leu Gly Ser Gly Met Lys225 230 235 240Leu Asn Asn Ser Cys Thr Pro Ile Thr Thr Pro Glu Leu Thr Thr Pro 245 250 255Cys Gly Ser Ala Glu Tyr Met Ala Pro Glu Val Val Glu Val Phe Thr 260 265 270Asp Gln Ala Thr Phe Tyr Asp Lys Arg Cys Asp Leu Trp Ser Leu Gly 275 280 285Val Val Leu Tyr Ile Met Leu Ser Gly Tyr Pro Pro Phe Val Gly His 290 295 300Cys Gly Ala Asp Cys Gly Trp Asp Arg Gly Glu Val Cys Arg Val Cys305 310 315 320Gln Asn Lys Leu Phe Glu Ser Ile Gln Glu Gly Lys Tyr Glu Phe Pro 325 330 335Asp Lys Asp Trp Ala His Ile Ser Ser Glu Ala Lys Asp Leu Ile Ser 340 345 350Lys Leu Leu Val Arg Asp Ala Lys Gln Arg Leu Ser Ala Ala Gln Val 355 360 365Leu Gln His Pro Trp Val Gln Gly Gln Ala Pro Glu Lys Gly Leu Pro 370 375 380Thr Pro Gln Val Leu Gln Arg Asn Ser Ser Thr Met Asp Leu Thr Leu385 390 395 400Phe Ala Ala Glu Ala Ile Ala Leu Asn Arg Gln Leu Ser Gln His Glu 405 410 415Glu Asn Glu Leu Ala Glu Glu Pro Glu Ala Leu Ala Asp Gly Leu Cys 420 425 430Ser Met Lys Leu Ser Pro Pro Cys Lys Ser Arg Leu Ala Arg Arg Arg 435 440 445Ala Leu Ala Gln Ala Gly Arg Gly Glu Asp Arg Ser Pro Pro Thr Ala 450 455 460Leu46534473PRTHomo sapiens 34Met Leu Lys Val Thr Val Pro Ser Cys Ser Ala Ser Ser Cys Ser Ser1 5 10 15Val Thr Ala Ser Ala Ala Pro Gly Thr Ala Ser Leu Val Pro Asp Tyr 20 25 30Trp Ile Asp Gly Ser Asn Arg Asp Ala Leu Ser Asp Phe Phe Glu Val 35 40 45Glu Ser Glu Leu Gly Arg Gly Ala Thr Ser Ile Val Tyr Arg Cys Lys 50 55 60Gln Lys Gly Thr Gln Lys Pro Tyr Ala Leu Lys Val Leu Lys Lys Thr65 70 75 80Val Asp Lys Lys Ile Val Arg Thr Glu Ile Gly Val Leu Leu Arg Leu 85 90 95Ser His Pro Asn Ile Ile Lys Leu Lys Glu Ile Phe Glu Thr Pro Thr 100 105 110Glu Ile Ser Leu Val Leu Glu Leu Val Thr Gly Gly Glu Leu Phe Asp 115 120 125Arg Ile Val Glu Lys Gly Tyr Tyr Ser Glu Arg Asp Ala Ala Asp Ala 130 135 140Val Lys Gln Ile Leu Glu Ala Val Ala Tyr Leu His Glu Asn Gly Ile145 150 155 160Val His Arg Asp Leu Lys Pro Glu Asn Leu Leu Tyr Ala Thr Pro Ala 165 170 175Pro Asp Ala Pro Leu Lys Ile Ala Asp Phe Gly Leu Ser Lys Ile Val 180 185 190Glu His Gln Val Leu Met Lys Thr Val Cys Gly Thr Pro Gly Tyr Cys 195 200 205Ala Pro Glu Ile Leu Arg Gly Cys Ala Tyr Gly Pro Glu Val Asp Met 210 215 220Trp Ser Val Gly Ile Ile Thr Tyr Ile Leu Leu Cys Gly Phe Glu Pro225 230 235 240Phe Tyr Asp Glu Arg Gly Asp Gln Phe Met Phe Arg Arg Ile Leu Asn 245 250 255Cys Glu Tyr Tyr Phe Ile Ser Pro Trp Trp Asp Glu Val Ser Leu Asn 260 265 270Ala Lys Asp Leu Val Arg Lys Leu Ile Val Leu Asp Pro Lys Lys Arg 275 280 285Leu Thr Thr Phe Gln Ala Leu Gln His Pro Trp Val Thr Gly Lys Ala 290 295 300Ala Asn Phe Val His Met Asp Thr Ala Gln Lys Lys Leu Gln Glu Phe305 310 315 320Asn Ala Arg Arg Lys Leu Lys Ala Ala Val Lys Ala Val Val Ala Ser 325 330 335Ser Arg Leu Gly Ser Ala Ser Ser Ser His Gly Ser Ile Gln Glu Ser 340 345 350His Lys Ala Ser Arg Asp Pro Ser Pro Ile Gln Asp Gly Asn Glu Asp 355 360 365Met Lys Ala Ile Pro Glu Gly Glu Lys Ile Gln Gly Asp Gly Ala Gln 370 375 380Ala Ala Val Lys Gly Ala Gln Ala Glu Leu Met Lys Val Gln Ala Leu385 390 395 400Glu Lys Val Lys Gly Ala Asp Ile Asn Ala Glu Glu Ala Pro Lys Met 405 410 415Val Pro Lys Ala Val Glu Asp Gly Ile Lys Val Ala Asp Leu Glu Leu 420 425 430Glu Glu Gly Leu Ala Glu Glu Lys Leu Lys Thr Val Glu Glu Ala Ala 435 440 445Ala Pro Arg Glu Gly Gln Gly Ser Ser Ala Val Gly Phe Glu Val Pro 450 455 460Gln Gln Asp Val Ile Leu Pro Glu Tyr465 47035367PRTHomo sapiens 35Met Asp Lys Glu Tyr Val Gly Phe Ala Ala Leu Pro Asn Gln Leu His1 5 10 15Arg Lys Ser Val Lys Lys Gly Phe Asp Phe Thr Leu Met Val Ala Gly 20 25 30Glu Ser Gly Leu Gly Lys Ser Thr Leu Ile Asn Ser Leu Phe Leu Thr 35 40 45Asn Leu Tyr Glu Asp Arg Gln Val Pro Glu Ala Ser Ala Arg Leu Thr 50 55 60Gln Thr Leu Ala Ile Glu Arg Arg Gly Val Glu Ile Glu Glu Gly Gly65 70 75 80Val Lys Val Lys Leu Thr Leu Val Asp Thr Pro Gly Phe Gly Asp Ser 85 90 95Val Asp Cys Ser Asp Cys Trp Leu Pro Val Val Lys Phe Ile Glu Glu 100 105 110Gln Phe Glu Gln Tyr Leu Arg Asp Glu Ser Gly Leu Asn Arg Lys Asn 115 120 125Ile Gln Asp Ser Arg Val His Cys Cys Leu Tyr Phe Ile Ser Pro Phe 130 135 140Gly Arg Gly Leu Arg Pro Leu Asp Val Ala Phe Leu Arg Ala Val His145 150 155 160Glu Lys Val Asn Ile Ile Pro Val Ile Gly Lys Ala Asp Ala Leu Met 165 170 175Pro Gln Glu Thr Gln Ala Leu Lys Gln Lys Ile Arg Asp Gln Leu Lys 180 185 190Glu Glu Glu Ile His Ile Tyr Gln Phe Pro Glu Cys Asp Ser Asp Glu 195 200 205Asp Glu Asp Phe Lys Arg Gln Asp Ala Glu Met Lys Glu Ser Ile Pro 210 215 220Phe Ala Val Val Gly Ser Cys Glu Val Val Arg Asp Gly Gly Asn Arg225 230 235 240Pro Val Arg Gly Arg Arg Tyr Ser Trp Gly Thr Val Glu Val Glu Asn 245 250 255Pro His His Cys Asp Phe Leu Asn Leu Arg Arg Met Leu Val Gln Thr 260 265 270His Leu Gln Asp Leu Lys Glu Val Thr His Asp Leu Leu Tyr Glu Gly 275 280 285Tyr Arg Ala Arg Cys Leu Gln Ser Leu Ala Arg Pro Gly Ala Arg Asp 290 295 300Arg Ala Ser Arg Ser Lys Leu Ser Arg Gln Ser Ala Thr Glu Ile Pro305 310 315 320Leu Pro Met Leu Pro Leu Ala Asp Thr Glu Lys Leu Ile Arg Glu Lys 325 330 335Asp Glu Glu Leu Arg Arg Met Gln Glu Met Leu Glu Lys Met Gln Ala 340 345 350Gln Met Gln Gln Ser Gln Ala Gln Gly Glu Gln Ser Asp Ala Leu 355 360 36536209PRTHomo sapiens 36Met Glu Gln Pro Arg Lys Ala Val Val Val Thr Gly Phe Gly Pro Phe1 5 10 15Gly Glu His Thr Val Asn Ala Ser Trp Ile Ala Val Gln Glu Leu Glu 20 25 30Lys Leu Gly Leu Gly Asp Ser Val Asp Leu His Val Tyr Glu Ile Pro 35 40 45Val Glu Tyr Gln Thr Val Gln Arg Leu Ile Pro Ala Leu Trp Glu Lys 50 55 60His Ser Pro Gln Leu Val Val His Val Gly Val Ser Gly Met Ala Thr65 70 75 80Thr Val Thr Leu Glu Lys Cys Gly His Asn Lys Gly Tyr Lys Gly Leu 85 90 95Asp Asn Cys Arg Phe Cys Pro Gly Ser Gln Cys Cys Val Glu Asp Gly 100 105 110Pro Glu Ser Ile Asp Ser Ile Ile Asp Met Asp Ala Val Cys Lys Arg 115 120 125Val Thr Thr Leu Gly Leu Asp Val Ser Val Thr Ile Ser Gln Asp Ala 130 135 140Gly Arg Lys Lys Pro Phe Pro Ala Lys Gly Asp Cys Val Phe Cys Arg145 150 155 160Arg Arg Arg Ala Arg Ser Leu Gln Ala Gln Cys Gly Phe Ser Leu Thr 165 170 175Pro Ala Leu Glu Leu Leu Pro Val Pro Phe Leu Lys Leu Leu Cys Pro
180 185 190Gly Pro Pro Arg Arg Arg Arg Ile Cys Arg Ile Leu Pro Gly Ala Gly 195 200 205Leu 37209PRTHomo sapiens 37Met Glu Gln Pro Arg Lys Ala Val Val Val Thr Gly Phe Gly Pro Phe1 5 10 15Gly Glu His Thr Val Asn Ala Ser Trp Ile Ala Val Gln Glu Leu Glu 20 25 30Lys Leu Gly Leu Gly Asp Ser Val Asp Leu His Val Tyr Glu Ile Pro 35 40 45Val Glu Tyr Gln Thr Val Gln Arg Leu Ile Pro Ala Leu Trp Glu Lys 50 55 60His Ser Pro Gln Leu Val Val His Val Gly Val Ser Gly Met Ala Thr65 70 75 80Thr Val Thr Leu Glu Lys Cys Gly His Asn Lys Gly Tyr Lys Gly Leu 85 90 95Asp Asn Cys Arg Phe Cys Pro Gly Ser Gln Cys Cys Val Glu Asp Gly 100 105 110Pro Glu Ser Ile Asp Ser Ile Ile Asp Met Asp Ala Val Cys Lys Arg 115 120 125Val Thr Thr Leu Gly Leu Asp Val Ser Val Thr Ile Ser Gln Asp Ala 130 135 140Gly Arg Tyr Leu Cys Asp Phe Thr Tyr Tyr Thr Ser Leu Tyr Gln Ser145 150 155 160His Gly Arg Ser Ala Phe Val His Val Pro Pro Leu Gly Lys Pro Tyr 165 170 175Asn Ala Asp Gln Leu Gly Arg Ala Leu Arg Ala Ile Ile Glu Glu Met 180 185 190Leu Asp Leu Leu Glu Gln Ser Glu Gly Lys Ile Asn Tyr Cys His Lys 195 200 205His 38359PRTHomo sapiens 38Met Ala Glu Ala Ile Thr Tyr Ala Asp Leu Arg Phe Val Lys Ala Pro1 5 10 15Leu Lys Lys Ser Ile Ser Ser Arg Leu Gly Gln Asp Pro Gly Ala Asp 20 25 30Asp Asp Gly Glu Ile Thr Tyr Glu Asn Val Gln Val Pro Ala Val Leu 35 40 45Gly Val Pro Ser Ser Leu Ala Ser Ser Val Leu Gly Asp Lys Ala Ala 50 55 60Val Lys Ser Glu Gln Pro Thr Ala Ser Trp Arg Ala Val Thr Ser Pro65 70 75 80Ala Val Gly Arg Ile Leu Pro Cys Arg Thr Thr Cys Leu Arg Tyr Leu 85 90 95Leu Leu Gly Leu Leu Leu Thr Cys Leu Leu Leu Gly Val Thr Ala Ile 100 105 110Cys Leu Gly Val Arg Tyr Leu Gln Val Ser Gln Gln Leu Gln Gln Thr 115 120 125Asn Arg Val Leu Glu Val Thr Asn Ser Ser Leu Arg Gln Gln Leu Arg 130 135 140Leu Lys Ile Thr Gln Leu Gly Gln Ser Ala Glu Asp Leu Gln Gly Ser145 150 155 160Arg Arg Glu Leu Ala Gln Ser Gln Glu Ala Leu Gln Val Glu Gln Arg 165 170 175Ala His Gln Ala Ala Glu Gly Gln Leu Gln Ala Cys Gln Ala Asp Arg 180 185 190Gln Lys Thr Lys Glu Thr Leu Gln Ser Glu Glu Gln Gln Arg Arg Ala 195 200 205Leu Glu Gln Lys Leu Ser Asn Met Glu Asn Arg Leu Lys Pro Phe Phe 210 215 220Thr Cys Gly Ser Ala Asp Thr Cys Cys Pro Ser Gly Trp Ile Met His225 230 235 240Gln Lys Ser Cys Phe Tyr Ile Ser Leu Thr Ser Lys Asn Trp Gln Glu 245 250 255Ser Gln Lys Gln Cys Glu Thr Leu Ser Ser Lys Leu Ala Thr Phe Ser 260 265 270Glu Ile Tyr Pro Gln Ser His Ser Tyr Tyr Phe Leu Asn Ser Leu Leu 275 280 285Pro Asn Gly Gly Ser Gly Asn Ser Tyr Trp Thr Gly Leu Ser Ser Asn 290 295 300Lys Asp Trp Lys Leu Thr Asp Asp Thr Gln Arg Thr Arg Thr Tyr Ala305 310 315 320Gln Ser Ser Lys Cys Asn Lys Val His Lys Thr Trp Ser Trp Trp Thr 325 330 335Leu Glu Ser Glu Ser Cys Arg Ser Ser Leu Pro Tyr Ile Cys Glu Met 340 345 350Thr Ala Phe Arg Phe Pro Asp 35539370PRTHomo sapiens 39Met Val Gly Lys Leu Lys Gln Asn Leu Leu Leu Ala Cys Leu Val Ile1 5 10 15Ser Ser Val Thr Val Phe Tyr Leu Gly Gln His Ala Met Glu Cys His 20 25 30His Arg Ile Glu Glu Arg Ser Gln Pro Val Lys Leu Glu Ser Thr Arg 35 40 45Thr Thr Val Arg Thr Gly Leu Asp Leu Lys Ala Asn Lys Thr Phe Ala 50 55 60Tyr His Lys Asp Met Pro Leu Ile Phe Ile Gly Gly Val Pro Arg Ser65 70 75 80Gly Thr Thr Leu Met Arg Ala Met Leu Asp Ala His Pro Asp Ile Arg 85 90 95Cys Gly Glu Glu Thr Arg Val Ile Pro Arg Ile Leu Ala Leu Lys Gln 100 105 110Met Trp Ser Arg Ser Ser Lys Glu Lys Ile Arg Leu Asp Glu Ala Gly 115 120 125Val Thr Asp Glu Val Leu Asp Ser Ala Met Gln Ala Phe Leu Leu Glu 130 135 140Ile Ile Val Lys His Gly Glu Pro Ala Pro Tyr Leu Cys Asn Lys Asp145 150 155 160Pro Phe Ala Leu Lys Ser Leu Thr Tyr Leu Ser Arg Leu Phe Pro Asn 165 170 175Ala Lys Phe Leu Leu Met Val Arg Asp Gly Arg Ala Ser Val His Ser 180 185 190Met Ile Ser Arg Lys Val Thr Ile Ala Gly Phe Asp Leu Asn Ser Tyr 195 200 205Arg Asp Cys Leu Thr Lys Trp Asn Arg Ala Ile Glu Thr Met Tyr Asn 210 215 220Gln Cys Met Glu Val Gly Tyr Lys Lys Cys Met Leu Val His Tyr Glu225 230 235 240Gln Leu Val Leu His Pro Glu Arg Trp Met Arg Thr Leu Leu Lys Phe 245 250 255Leu Gln Ile Pro Trp Asn His Ser Val Leu His His Glu Glu Met Ile 260 265 270Gly Lys Ala Gly Gly Val Ser Leu Ser Lys Val Glu Arg Ser Thr Asp 275 280 285Gln Val Ile Lys Pro Val Asn Val Gly Ala Leu Ser Lys Trp Val Gly 290 295 300Lys Ile Pro Pro Asp Val Leu Gln Asp Met Ala Val Ile Ala Pro Met305 310 315 320Leu Ala Lys Leu Gly Tyr Asp Pro Tyr Ala Asn Pro Pro Asn Tyr Gly 325 330 335Lys Pro Asp Pro Lys Ile Ile Glu Asn Thr Arg Arg Val Tyr Lys Gly 340 345 350Glu Phe Gln Leu Pro Asp Phe Leu Lys Glu Lys Pro Gln Thr Glu Gln 355 360 365Val Glu 37040349PRTHomo sapiens 40Met Asn Ser Thr Leu Asp Gly Asn Gln Ser Ser His Pro Phe Cys Leu1 5 10 15Leu Ala Phe Gly Tyr Leu Glu Thr Val Asn Phe Cys Leu Leu Glu Val 20 25 30Leu Ile Ile Val Phe Leu Thr Val Leu Ile Ile Ser Gly Asn Ile Ile 35 40 45Val Ile Phe Val Phe His Cys Ala Pro Leu Leu Asn His His Thr Thr 50 55 60Ser Tyr Phe Ile Gln Thr Met Ala Tyr Ala Asp Leu Phe Val Gly Val65 70 75 80Ser Cys Val Val Pro Ser Leu Ser Leu Leu His His Pro Leu Pro Val 85 90 95Glu Glu Ser Leu Thr Cys Gln Ile Phe Gly Phe Val Val Ser Val Leu 100 105 110Lys Ser Val Ser Met Ala Ser Leu Ala Cys Ile Ser Ile Asp Arg Tyr 115 120 125Ile Ala Ile Thr Lys Pro Leu Thr Tyr Asn Thr Leu Val Thr Pro Trp 130 135 140Arg Leu Arg Leu Cys Ile Phe Leu Ile Trp Leu Tyr Ser Thr Leu Val145 150 155 160Phe Leu Pro Ser Phe Phe His Trp Gly Lys Pro Gly Tyr His Gly Asp 165 170 175Val Phe Gln Trp Cys Ala Glu Ser Trp His Thr Asp Ser Tyr Phe Thr 180 185 190Leu Phe Ile Val Met Met Leu Tyr Ala Pro Ala Ala Leu Ile Val Cys 195 200 205Phe Thr Tyr Phe Asn Ile Phe Arg Ile Cys Gln Gln His Thr Lys Asp 210 215 220Ile Ser Glu Arg Gln Ala Arg Phe Ser Ser Gln Ser Gly Glu Thr Gly225 230 235 240Glu Val Gln Ala Cys Pro Asp Lys Arg Tyr Ala Met Val Leu Phe Arg 245 250 255Ile Thr Ser Val Phe Tyr Ile Leu Trp Leu Pro Tyr Ile Ile Tyr Phe 260 265 270Leu Leu Glu Ser Ser Thr Gly His Ser Asn Arg Phe Ala Ser Phe Leu 275 280 285Thr Thr Trp Leu Ala Ile Ser Asn Ser Phe Cys Asn Cys Val Ile Tyr 290 295 300Ser Leu Ser Asn Ser Val Phe Gln Arg Gly Leu Lys Arg Leu Ser Gly305 310 315 320Ala Met Cys Thr Ser Cys Ala Ser Gln Thr Thr Ala Asn Asp Pro Tyr 325 330 335Thr Val Arg Ser Lys Gly Pro Leu Asn Gly Cys His Ile 340 34541565PRTHomo sapiens 41Met Pro Gln Ala Ser Glu His Arg Leu Gly Arg Thr Arg Glu Pro Pro1 5 10 15Val Asn Ile Gln Pro Arg Val Gly Ser Lys Leu Pro Phe Ala Pro Arg 20 25 30Ala Arg Ser Lys Glu Arg Arg Asn Pro Ala Ser Gly Pro Asn Pro Met 35 40 45Leu Arg Pro Leu Pro Pro Arg Pro Gly Leu Pro Asp Glu Arg Leu Lys 50 55 60Lys Leu Glu Leu Gly Arg Gly Arg Thr Ser Gly Pro Arg Pro Arg Gly65 70 75 80Pro Leu Arg Ala Asp His Gly Val Pro Leu Pro Gly Ser Pro Pro Pro 85 90 95Thr Val Ala Leu Pro Leu Pro Ser Arg Thr Asn Leu Ala Arg Ser Lys 100 105 110Ser Val Ser Ser Gly Asp Leu Arg Pro Met Gly Ile Ala Leu Gly Gly 115 120 125His Arg Gly Thr Gly Glu Leu Gly Ala Ala Leu Ser Arg Leu Ala Leu 130 135 140Arg Pro Glu Pro Pro Thr Leu Arg Arg Ser Thr Ser Leu Arg Arg Leu145 150 155 160Gly Gly Phe Pro Gly Pro Pro Thr Leu Phe Ser Ile Arg Thr Glu Pro 165 170 175Pro Ala Ser His Gly Ser Phe His Met Ile Ser Ala Arg Ser Ser Glu 180 185 190Pro Phe Tyr Ser Asp Asp Lys Met Ala His His Thr Leu Leu Leu Gly 195 200 205Ser Gly His Val Gly Leu Arg Asn Leu Gly Asn Thr Cys Phe Leu Asn 210 215 220Ala Val Leu Gln Cys Leu Ser Ser Thr Arg Pro Leu Arg Asp Phe Cys225 230 235 240Leu Arg Arg Asp Phe Arg Gln Glu Val Pro Gly Gly Gly Arg Ala Gln 245 250 255Glu Leu Thr Glu Ala Phe Ala Asp Val Ile Gly Ala Leu Trp His Pro 260 265 270Asp Ser Cys Glu Ala Val Asn Pro Thr Arg Phe Arg Ala Val Phe Gln 275 280 285Lys Tyr Val Pro Ser Phe Ser Gly Tyr Ser Gln Gln Asp Ala Gln Glu 290 295 300Phe Leu Lys Leu Leu Met Glu Arg Leu His Leu Glu Ile Asn Arg Arg305 310 315 320Gly Arg Arg Ala Pro Pro Ile Leu Ala Asn Gly Pro Val Pro Ser Pro 325 330 335Pro Arg Arg Gly Gly Ala Leu Leu Glu Glu Pro Glu Leu Ser Asp Asp 340 345 350Asp Arg Ala Asn Leu Met Trp Lys Arg Tyr Leu Glu Arg Glu Asp Ser 355 360 365Lys Ile Val Asp Leu Phe Val Gly Gln Leu Lys Ser Cys Leu Lys Cys 370 375 380Gln Ala Cys Gly Tyr Arg Ser Thr Thr Phe Glu Val Phe Cys Asp Leu385 390 395 400Ser Leu Pro Ile Pro Lys Lys Gly Phe Ala Gly Gly Lys Val Ser Leu 405 410 415Arg Asp Cys Phe Asn Leu Phe Thr Lys Glu Glu Glu Leu Glu Ser Glu 420 425 430Asn Ala Pro Val Cys Asp Arg Cys Arg Gln Lys Thr Arg Ser Thr Lys 435 440 445Lys Leu Thr Val Gln Arg Phe Pro Arg Ile Leu Gly Leu Asp Leu Asn 450 455 460Arg Phe Ser Ala Ser Arg Gly Ser Ile Lys Lys Ser Ser Val Gly Val465 470 475 480Asp Phe Pro Leu Gln Arg Leu Ser Leu Gly Asp Phe Ala Ser Asp Lys 485 490 495Ala Gly Ser Pro Val Tyr Gln Leu Tyr Ala Leu Cys Asn His Ser Gly 500 505 510Ser Val His Tyr Gly His Tyr Thr Ala Leu Cys Arg Cys Gln Thr Gly 515 520 525Trp His Val Tyr Asn Asp Ser Arg Val Ser Pro Val Ser Glu Asn Gln 530 535 540Val Ala Ser Ser Glu Gly Tyr Val Leu Phe Tyr Gln Leu Met Gln Glu545 550 555 560Pro Pro Arg Cys Leu 56542551PRTHomo sapiens 42Met Pro Gln Ala Ser Glu His Arg Leu Gly Arg Thr Arg Glu Pro Pro1 5 10 15Val Asn Ile Gln Pro Arg Val Gly Ser Lys Leu Pro Phe Ala Pro Arg 20 25 30Ala Arg Ser Lys Glu Arg Arg Asn Pro Ala Ser Gly Pro Asn Pro Met 35 40 45Leu Arg Pro Leu Pro Pro Arg Pro Gly Leu Pro Asp Glu Arg Leu Lys 50 55 60Lys Leu Glu Leu Gly Arg Gly Arg Thr Ser Gly Pro Arg Pro Arg Gly65 70 75 80Pro Leu Arg Ala Asp His Gly Val Pro Leu Pro Gly Ser Pro Pro Pro 85 90 95Thr Val Ala Leu Pro Leu Pro Ser Arg Thr Asn Leu Ala Arg Ser Lys 100 105 110Ser Val Ser Ser Gly Asp Leu Arg Pro Met Gly Ile Ala Leu Gly Gly 115 120 125His Arg Gly Thr Gly Glu Leu Gly Ala Ala Leu Ser Arg Leu Ala Leu 130 135 140Arg Pro Glu Pro Pro Thr Leu Arg Arg Ser Thr Ser Leu Arg Arg Leu145 150 155 160Gly Gly Phe Pro Gly Pro Pro Thr Leu Phe Ser Ile Arg Thr Glu Pro 165 170 175Pro Ala Ser His Gly Ser Phe His Met Ile Ser Ala Arg Ser Ser Glu 180 185 190Pro Phe Tyr Ser Asp Asp Lys Met Ala His His Thr Leu Leu Leu Gly 195 200 205Ser Gly His Val Gly Leu Arg Asn Leu Gly Asn Thr Cys Phe Leu Asn 210 215 220Ala Val Leu Gln Cys Leu Ser Ser Thr Arg Pro Leu Arg Asp Phe Cys225 230 235 240Leu Arg Arg Asp Phe Arg Gln Glu Val Pro Gly Gly Gly Arg Ala Gln 245 250 255Glu Leu Thr Glu Ala Phe Ala Asp Val Ile Gly Ala Leu Trp His Pro 260 265 270Asp Ser Cys Glu Ala Val Asn Pro Thr Arg Phe Arg Ala Val Phe Gln 275 280 285Lys Tyr Val Pro Ser Phe Ser Gly Tyr Ser Gln Gln Asp Ala Gln Glu 290 295 300Phe Leu Lys Leu Leu Met Glu Arg Leu His Leu Glu Ile Asn Arg Arg305 310 315 320Gly Arg Arg Ala Pro Pro Ile Leu Ala Asn Gly Pro Val Pro Ser Pro 325 330 335Pro Arg Arg Gly Gly Ala Leu Leu Glu Glu Pro Glu Leu Ser Asp Asp 340 345 350Asp Arg Ala Asn Leu Met Trp Lys Arg Tyr Leu Glu Arg Glu Asp Ser 355 360 365Lys Ile Val Asp Leu Phe Val Gly Gln Leu Lys Ser Cys Leu Lys Cys 370 375 380Gln Ala Cys Gly Tyr Arg Ser Thr Thr Phe Glu Val Phe Cys Asp Leu385 390 395 400Ser Leu Pro Ile Pro Lys Lys Gly Phe Ala Gly Gly Lys Val Ser Leu 405 410 415Arg Asp Cys Phe Asn Leu Phe Thr Lys Glu Glu Glu Leu Glu Ser Glu 420 425 430Asn Ala Pro Val Cys Asp Arg Cys Arg Gln Lys Thr Arg Ser Thr Lys 435 440 445Lys Leu Thr Val Gln Arg Phe Pro Arg Ile Leu Gly Leu Asp Leu Asn 450 455 460Arg Phe Ser Ala Ser Arg Gly Ser Ile Lys Lys Ser Ser Val Gly Val465 470 475 480Asp Phe Pro Leu Gln Arg Leu Ser Leu Gly Asp Phe Ala Ser Asp Lys 485 490 495Ala Gly Ser Val His Tyr Gly His Tyr Thr Ala Leu Cys Arg Cys Gln 500 505 510Thr Gly Trp His Val Tyr Asn Asp Ser Arg Val Ser Pro Val Ser Glu 515 520 525Asn Gln Val Ala Ser Ser Glu Gly Tyr Val Leu Phe Tyr Gln Leu Met 530 535 540Gln Glu Pro Pro Arg Cys Leu545 55043537PRTHomo sapiens 43Met Ala Trp Arg Gly Ala Gly Pro Ser Val Pro Gly Ala Pro Gly Gly1 5 10 15Val Gly Leu Ser Leu Gly Leu Leu Leu Gln Leu Leu Leu Leu Leu Gly 20 25
30Pro Ala Arg Gly Phe Gly Asp Glu Glu Glu Arg Arg Cys Asp Pro Ile 35 40 45Arg Ile Ser Met Cys Gln Asn Leu Gly Tyr Asn Val Thr Lys Met Pro 50 55 60Asn Leu Val Gly His Glu Leu Gln Thr Asp Ala Glu Leu Gln Leu Thr65 70 75 80Thr Phe Thr Pro Leu Ile Gln Tyr Gly Cys Ser Ser Gln Leu Gln Phe 85 90 95Phe Leu Cys Ser Val Tyr Val Pro Met Cys Thr Glu Lys Ile Asn Ile 100 105 110Pro Ile Gly Pro Cys Gly Gly Met Cys Leu Ser Val Lys Arg Arg Cys 115 120 125Glu Pro Val Leu Lys Glu Phe Gly Phe Ala Trp Pro Glu Ser Leu Asn 130 135 140Cys Ser Lys Phe Pro Pro Gln Asn Asp His Asn His Met Cys Met Glu145 150 155 160Gly Pro Gly Asp Glu Glu Val Pro Leu Pro His Lys Thr Pro Ile Gln 165 170 175Pro Gly Glu Glu Cys His Ser Val Gly Thr Asn Ser Asp Gln Tyr Ile 180 185 190Trp Val Lys Arg Ser Leu Asn Cys Val Leu Lys Cys Gly Tyr Asp Ala 195 200 205Gly Leu Tyr Ser Arg Ser Ala Lys Glu Phe Thr Asp Ile Trp Met Ala 210 215 220Val Trp Ala Ser Leu Cys Phe Ile Ser Thr Ala Phe Thr Val Leu Thr225 230 235 240Phe Leu Ile Asp Ser Ser Arg Phe Ser Tyr Pro Glu Arg Pro Ile Ile 245 250 255Phe Leu Ser Met Cys Tyr Asn Ile Tyr Ser Ile Ala Tyr Ile Val Arg 260 265 270Leu Thr Val Gly Arg Glu Arg Ile Ser Cys Asp Phe Glu Glu Ala Ala 275 280 285Glu Pro Val Leu Ile Gln Glu Gly Leu Lys Asn Thr Gly Cys Ala Ile 290 295 300Ile Phe Leu Leu Met Tyr Phe Phe Gly Met Ala Ser Ser Ile Trp Trp305 310 315 320Val Ile Leu Thr Leu Thr Trp Phe Leu Ala Ala Gly Leu Lys Trp Gly 325 330 335His Glu Ala Ile Glu Met His Ser Ser Tyr Phe His Ile Ala Ala Trp 340 345 350Ala Ile Pro Ala Val Lys Thr Ile Val Ile Leu Ile Met Arg Leu Val 355 360 365Asp Ala Asp Glu Leu Thr Gly Leu Cys Tyr Val Gly Asn Gln Asn Leu 370 375 380Asp Ala Leu Thr Gly Phe Val Val Ala Pro Leu Phe Thr Tyr Leu Val385 390 395 400Ile Gly Thr Leu Phe Ile Ala Ala Gly Leu Val Ala Leu Phe Lys Ile 405 410 415Arg Ser Asn Leu Gln Lys Asp Gly Thr Lys Thr Asp Lys Leu Glu Arg 420 425 430Leu Met Val Lys Ile Gly Val Phe Ser Val Leu Tyr Thr Val Pro Ala 435 440 445Thr Cys Val Ile Ala Cys Tyr Phe Tyr Glu Ile Ser Asn Trp Ala Leu 450 455 460Phe Arg Tyr Ser Ala Asp Asp Ser Asn Met Ala Val Glu Met Leu Lys465 470 475 480Ile Phe Met Ser Leu Leu Val Gly Ile Thr Ser Gly Met Trp Ile Trp 485 490 495Ser Ala Lys Thr Leu His Thr Trp Gln Lys Cys Ser Asn Arg Leu Val 500 505 510Asn Ser Gly Lys Val Lys Arg Glu Lys Arg Gly Asn Gly Trp Val Lys 515 520 525Pro Gly Lys Gly Ser Glu Thr Val Val 530 53544433PRTHomo sapiens 44Met Cys Thr Glu Lys Ile Asn Ile Pro Ile Gly Pro Cys Gly Gly Met1 5 10 15Cys Leu Ser Val Lys Arg Arg Cys Glu Pro Val Leu Lys Glu Phe Gly 20 25 30Phe Ala Trp Pro Glu Ser Leu Asn Cys Ser Lys Phe Pro Pro Gln Asn 35 40 45Asp His Asn His Met Cys Met Glu Gly Pro Gly Asp Glu Glu Val Pro 50 55 60Leu Pro His Lys Thr Pro Ile Gln Pro Gly Glu Glu Cys His Ser Val65 70 75 80Gly Thr Asn Ser Asp Gln Tyr Ile Trp Val Lys Arg Ser Leu Asn Cys 85 90 95Val Leu Lys Cys Gly Tyr Asp Ala Gly Leu Tyr Ser Arg Ser Ala Lys 100 105 110Glu Phe Thr Asp Ile Trp Met Ala Val Trp Ala Ser Leu Cys Phe Ile 115 120 125Ser Thr Ala Phe Thr Val Leu Thr Phe Leu Ile Asp Ser Ser Arg Phe 130 135 140Ser Tyr Pro Glu Arg Pro Ile Ile Phe Leu Ser Met Cys Tyr Asn Ile145 150 155 160Tyr Ser Ile Ala Tyr Ile Val Arg Leu Thr Val Gly Arg Glu Arg Ile 165 170 175Ser Cys Asp Phe Glu Glu Ala Ala Glu Pro Val Leu Ile Gln Glu Gly 180 185 190Leu Lys Asn Thr Gly Cys Ala Ile Ile Phe Leu Leu Met Tyr Phe Phe 195 200 205Gly Met Ala Ser Ser Ile Trp Trp Val Ile Leu Thr Leu Thr Trp Phe 210 215 220Leu Ala Ala Gly Leu Lys Trp Gly His Glu Ala Ile Glu Met His Ser225 230 235 240Ser Tyr Phe His Ile Ala Ala Trp Ala Ile Pro Ala Val Lys Thr Ile 245 250 255Val Ile Leu Ile Met Arg Leu Val Asp Ala Asp Glu Leu Thr Gly Leu 260 265 270Cys Tyr Val Gly Asn Gln Asn Leu Asp Ala Leu Thr Gly Phe Val Val 275 280 285Ala Pro Leu Phe Thr Tyr Leu Val Ile Gly Thr Leu Phe Ile Ala Ala 290 295 300Gly Leu Val Ala Leu Phe Lys Ile Arg Ser Asn Leu Gln Lys Asp Gly305 310 315 320Thr Lys Thr Asp Lys Leu Glu Arg Leu Met Val Lys Ile Gly Val Phe 325 330 335Ser Val Leu Tyr Thr Val Pro Ala Thr Cys Val Ile Ala Cys Tyr Phe 340 345 350Tyr Glu Ile Ser Asn Trp Ala Leu Phe Arg Tyr Ser Ala Asp Asp Ser 355 360 365Asn Met Ala Val Glu Met Leu Lys Ile Phe Met Ser Leu Leu Val Gly 370 375 380Ile Thr Ser Gly Met Trp Ile Trp Ser Ala Lys Thr Leu His Thr Trp385 390 395 400Gln Lys Cys Ser Asn Arg Leu Val Asn Ser Gly Lys Val Lys Arg Glu 405 410 415Lys Arg Gly Asn Gly Trp Val Lys Pro Gly Lys Gly Ser Glu Thr Val 420 425 430Val45399PRTHomo sapiens 45Met Arg Pro Glu Arg Pro Arg Pro Arg Gly Ser Ala Pro Gly Pro Met1 5 10 15Glu Thr Pro Pro Trp Asp Pro Ala Arg Asn Asp Ser Leu Pro Pro Thr 20 25 30Leu Thr Pro Ala Val Pro Pro Tyr Val Lys Leu Gly Leu Thr Val Val 35 40 45Tyr Thr Val Phe Tyr Ala Leu Leu Phe Val Phe Ile Tyr Val Gln Leu 50 55 60Trp Leu Val Leu Arg Tyr Arg His Lys Arg Leu Ser Tyr Gln Ser Val65 70 75 80Phe Leu Phe Leu Cys Leu Phe Trp Ala Ser Leu Arg Thr Val Leu Phe 85 90 95Ser Phe Tyr Phe Lys Asp Phe Val Ala Ala Asn Ser Leu Ser Pro Phe 100 105 110Val Phe Trp Leu Leu Tyr Cys Phe Pro Val Cys Leu Gln Phe Phe Thr 115 120 125Leu Thr Leu Met Asn Leu Tyr Phe Thr Gln Val Ile Phe Lys Ala Lys 130 135 140Ser Lys Tyr Ser Pro Glu Leu Leu Lys Tyr Arg Leu Pro Leu Tyr Leu145 150 155 160Ala Ser Leu Phe Ile Ser Leu Val Phe Leu Leu Val Asn Leu Thr Cys 165 170 175Ala Val Leu Val Lys Thr Gly Asn Trp Glu Arg Lys Val Ile Val Ser 180 185 190Val Arg Val Ala Ile Asn Asp Thr Leu Phe Val Leu Cys Ala Val Ser 195 200 205Leu Ser Ile Cys Leu Tyr Lys Ile Ser Lys Met Ser Leu Ala Asn Ile 210 215 220Tyr Leu Glu Ser Lys Gly Ser Ser Val Cys Gln Val Thr Ala Ile Gly225 230 235 240Val Thr Val Ile Leu Leu Tyr Thr Ser Arg Ala Cys Tyr Asn Leu Phe 245 250 255Ile Leu Ser Phe Ser Gln Asn Lys Ser Val His Ser Phe Asp Tyr Asp 260 265 270Trp Tyr Asn Val Ser Asp Gln Ala Asp Leu Lys Asn Gln Leu Gly Asp 275 280 285Ala Gly Tyr Val Leu Phe Gly Val Val Leu Phe Val Trp Glu Leu Leu 290 295 300Pro Thr Thr Leu Val Val Tyr Phe Phe Arg Val Arg Asn Pro Thr Lys305 310 315 320Asp Leu Thr Asn Pro Gly Met Val Pro Ser His Gly Phe Ser Pro Arg 325 330 335Ser Tyr Phe Phe Asp Asn Pro Arg Arg Tyr Asp Ser Asp Asp Asp Leu 340 345 350Ala Trp Asn Ile Ala Pro Gln Gly Leu Gln Gly Gly Phe Ala Pro Asp 355 360 365Tyr Tyr Asp Trp Gly Gln Gln Thr Asn Ser Phe Leu Ala Gln Ala Gly 370 375 380Thr Leu Gln Asp Ser Thr Leu Asp Pro Asp Lys Pro Ser Leu Gly385 390 39546178PRTHomo sapiens 46Met Ser Pro Ser Gly Arg Leu Cys Leu Leu Thr Ile Val Gly Leu Ile1 5 10 15Leu Pro Thr Arg Gly Gln Thr Leu Lys Asp Thr Thr Ser Ser Ser Ser 20 25 30Ala Asp Ser Thr Ile Met Asp Ile Gln Val Pro Thr Arg Ala Pro Asp 35 40 45Ala Val Tyr Thr Glu Leu Gln Pro Thr Ser Pro Thr Pro Thr Trp Pro 50 55 60Ala Asp Glu Thr Pro Gln Pro Gln Thr Gln Thr Gln Gln Leu Glu Gly65 70 75 80Thr Asp Gly Pro Leu Val Thr Asp Pro Glu Thr His Lys Ser Thr Lys 85 90 95Ala Ala His Pro Thr Asp Asp Thr Thr Thr Leu Ser Glu Arg Pro Ser 100 105 110Pro Ser Thr Asp Val Gln Thr Asp Pro Gln Thr Leu Lys Pro Ser Gly 115 120 125Phe His Glu Asp Asp Pro Phe Phe Tyr Asp Glu His Thr Leu Arg Lys 130 135 140Arg Gly Leu Leu Val Ala Ala Val Leu Phe Ile Thr Gly Ile Ile Ile145 150 155 160Leu Thr Ser Gly Lys Cys Arg Gln Leu Ser Arg Leu Cys Arg Asn Arg 165 170 175Cys Arg47219PRTHomo sapiens 47Met Asp Ser Gly Thr Arg Pro Val Gly Ser Cys Cys Ser Ser Pro Ala1 5 10 15Gly Leu Ser Arg Glu Tyr Lys Leu Val Met Leu Gly Ala Gly Gly Val 20 25 30Gly Lys Ser Ala Met Thr Met Gln Phe Ile Ser His Arg Phe Pro Glu 35 40 45Asp His Asp Pro Thr Ile Glu Asp Ala Tyr Lys Ile Arg Ile Arg Ile 50 55 60Asp Asp Glu Pro Ala Asn Leu Asp Ile Leu Asp Thr Ala Gly Gln Ala65 70 75 80Glu Phe Thr Ala Met Arg Asp Gln Tyr Met Arg Ala Gly Glu Gly Phe 85 90 95Ile Ile Cys Tyr Ser Ile Thr Asp Arg Arg Ser Phe His Glu Val Arg 100 105 110Glu Phe Lys Gln Leu Ile Tyr Arg Val Arg Arg Thr Asp Asp Thr Pro 115 120 125Val Val Leu Val Gly Asn Lys Ser Asp Leu Lys Gln Leu Arg Gln Val 130 135 140Thr Lys Glu Glu Gly Leu Ala Leu Ala Arg Glu Phe Ser Cys Pro Phe145 150 155 160Phe Glu Thr Ser Ala Ala Tyr Arg Tyr Tyr Ile Asp Asp Val Phe His 165 170 175Ala Leu Val Arg Glu Ile Arg Arg Lys Glu Lys Glu Ala Val Leu Ala 180 185 190Met Glu Lys Lys Ser Lys Pro Lys Asn Ser Val Trp Lys Arg Leu Lys 195 200 205Ser Pro Phe Arg Lys Lys Lys Asp Ser Val Thr 210 21548479PRTHomo sapiens 48Met Lys Ser Gln Gly Gln His Trp Tyr Ser Ser Ser Asp Lys Asn Cys1 5 10 15Lys Val Ser Phe Arg Glu Lys Leu Leu Ile Ile Asp Ser Asn Leu Gly 20 25 30Val Gln Asp Val Glu Asn Leu Lys Phe Leu Cys Ile Gly Leu Val Pro 35 40 45Asn Lys Lys Leu Glu Lys Ser Ser Ser Ala Ser Asp Val Phe Glu His 50 55 60Leu Leu Ala Glu Asp Leu Leu Ser Glu Glu Asp Pro Phe Phe Leu Ala65 70 75 80Glu Leu Leu Tyr Ile Ile Arg Gln Lys Lys Leu Leu Gln His Leu Asn 85 90 95Cys Thr Lys Glu Glu Val Glu Arg Leu Leu Pro Thr Arg Gln Arg Val 100 105 110Ser Leu Phe Arg Asn Leu Leu Tyr Glu Leu Ser Glu Gly Ile Asp Ser 115 120 125Glu Asn Leu Lys Asp Met Ile Phe Leu Leu Lys Asp Ser Leu Pro Lys 130 135 140Thr Glu Met Thr Ser Leu Ser Phe Leu Ala Phe Leu Glu Lys Gln Gly145 150 155 160Lys Ile Asp Glu Asp Asn Leu Thr Cys Leu Glu Asp Leu Cys Lys Thr 165 170 175Val Val Pro Lys Leu Leu Arg Asn Ile Glu Lys Tyr Lys Arg Glu Lys 180 185 190Ala Ile Gln Ile Val Thr Pro Pro Val Asp Lys Glu Ala Glu Ser Tyr 195 200 205Gln Gly Glu Glu Glu Leu Val Ser Gln Thr Asp Val Lys Thr Phe Leu 210 215 220Glu Ala Leu Pro Arg Ala Ala Val Tyr Arg Met Asn Arg Asn His Arg225 230 235 240Gly Leu Cys Val Ile Val Asn Asn His Ser Phe Thr Ser Leu Lys Asp 245 250 255Arg Gln Gly Thr His Lys Asp Ala Glu Ile Leu Ser His Val Phe Gln 260 265 270Trp Leu Gly Phe Thr Val His Ile His Asn Asn Val Thr Lys Val Glu 275 280 285Met Glu Met Val Leu Gln Lys Gln Lys Cys Asn Pro Ala His Ala Asp 290 295 300Gly Asp Cys Phe Val Phe Cys Ile Leu Thr His Gly Arg Phe Gly Ala305 310 315 320Val Tyr Ser Ser Asp Glu Ala Leu Ile Pro Ile Arg Glu Ile Met Ser 325 330 335His Phe Thr Ala Leu Gln Cys Pro Arg Leu Ala Glu Lys Pro Lys Leu 340 345 350Phe Phe Ile Gln Ala Cys Gln Gly Glu Glu Ile Gln Pro Ser Val Ser 355 360 365Ile Glu Ala Asp Ala Leu Asn Pro Glu Gln Ala Pro Thr Ser Leu Gln 370 375 380Asp Ser Ile Pro Ala Glu Ala Asp Phe Leu Leu Gly Leu Ala Thr Val385 390 395 400Pro Gly Tyr Val Ser Phe Arg His Val Glu Glu Gly Ser Trp Tyr Ile 405 410 415Gln Ser Leu Cys Asn His Leu Lys Lys Leu Val Pro Arg His Glu Asp 420 425 430Ile Leu Ser Ile Leu Thr Ala Val Asn Asp Asp Val Ser Arg Arg Val 435 440 445Asp Lys Gln Gly Thr Lys Lys Gln Met Pro Gln Pro Ala Phe Thr Leu 450 455 460Arg Lys Lys Leu Val Phe Pro Val Pro Leu Asp Ala Leu Ser Leu465 470 47549521PRTHomo sapiens 49Met Lys Ser Gln Gly Gln His Trp Tyr Ser Ser Ser Asp Lys Asn Cys1 5 10 15Lys Val Ser Phe Arg Glu Lys Leu Leu Ile Ile Asp Ser Asn Leu Gly 20 25 30Val Gln Asp Val Glu Asn Leu Lys Phe Leu Cys Ile Gly Leu Val Pro 35 40 45Asn Lys Lys Leu Glu Lys Ser Ser Ser Ala Ser Asp Val Phe Glu His 50 55 60Leu Leu Ala Glu Asp Leu Leu Ser Glu Glu Asp Pro Phe Phe Leu Ala65 70 75 80Glu Leu Leu Tyr Ile Ile Arg Gln Lys Lys Leu Leu Gln His Leu Asn 85 90 95Cys Thr Lys Glu Glu Val Glu Arg Leu Leu Pro Thr Arg Gln Arg Val 100 105 110Ser Leu Phe Arg Asn Leu Leu Tyr Glu Leu Ser Glu Gly Ile Asp Ser 115 120 125Glu Asn Leu Lys Asp Met Ile Phe Leu Leu Lys Asp Ser Leu Pro Lys 130 135 140Thr Glu Met Thr Ser Leu Ser Phe Leu Ala Phe Leu Glu Lys Gln Gly145 150 155 160Lys Ile Asp Glu Asp Asn Leu Thr Cys Leu Glu Asp Leu Cys Lys Thr 165 170 175Val Val Pro Lys Leu Leu Arg Asn Ile Glu Lys Tyr Lys Arg Glu Lys 180 185 190Ala Ile Gln Ile Val Thr Pro Pro Val Asp Lys Glu Ala Glu Ser Tyr 195 200 205Gln Gly Glu Glu Glu Leu Val Ser Gln Thr Asp Val Lys Thr Phe Leu 210 215 220Glu Ala Leu Pro Gln Glu Ser Trp Gln Asn Lys His Ala Gly Ser Asn225 230 235 240Gly Asn Arg Ala Thr Asn Gly Ala Pro Ser Leu Val Ser Arg Gly Met 245
250 255Gln Gly Ala Ser Ala Asn Thr Leu Asn Ser Glu Thr Ser Thr Lys Arg 260 265 270Ala Ala Val Tyr Arg Met Asn Arg Asn His Arg Gly Leu Cys Val Ile 275 280 285Val Asn Asn His Ser Phe Thr Ser Leu Lys Asp Arg Gln Gly Thr His 290 295 300Lys Asp Ala Glu Ile Leu Ser His Val Phe Gln Trp Leu Gly Phe Thr305 310 315 320Val His Ile His Asn Asn Val Thr Lys Val Glu Met Glu Met Val Leu 325 330 335Gln Lys Gln Lys Cys Asn Pro Ala His Ala Asp Gly Asp Cys Phe Val 340 345 350Phe Cys Ile Leu Thr His Gly Arg Phe Gly Ala Val Tyr Ser Ser Asp 355 360 365Glu Ala Leu Ile Pro Ile Arg Glu Ile Met Ser His Phe Thr Ala Leu 370 375 380Gln Cys Pro Arg Leu Ala Glu Lys Pro Lys Leu Phe Phe Ile Gln Ala385 390 395 400Cys Gln Gly Glu Glu Ile Gln Pro Ser Val Ser Ile Glu Ala Asp Ala 405 410 415Leu Asn Pro Glu Gln Ala Pro Thr Ser Leu Gln Asp Ser Ile Pro Ala 420 425 430Glu Ala Asp Phe Leu Leu Gly Leu Ala Thr Val Pro Gly Tyr Val Ser 435 440 445Phe Arg His Val Glu Glu Gly Ser Trp Tyr Ile Gln Ser Leu Cys Asn 450 455 460His Leu Lys Lys Leu Val Pro Arg Met Leu Lys Phe Leu Glu Lys Thr465 470 475 480Met Glu Ile Arg Gly Arg Lys Arg Thr Val Trp Gly Ala Lys Gln Ile 485 490 495Ser Ala Thr Ser Leu Pro Thr Ala Ile Ser Ala Gln Thr Pro Arg Pro 500 505 510Pro Met Arg Arg Trp Ser Ser Val Ser 515 52050273PRTHomo sapiens 50Met Lys Ser Gln Gly Gln His Trp Tyr Ser Ser Ser Asp Lys Asn Cys1 5 10 15Lys Val Ser Phe Arg Glu Lys Leu Leu Ile Ile Asp Ser Asn Leu Gly 20 25 30Val Gln Asp Val Glu Asn Leu Lys Phe Leu Cys Ile Gly Leu Val Pro 35 40 45Asn Lys Lys Leu Glu Lys Ser Ser Ser Ala Ser Asp Val Phe Glu His 50 55 60Leu Leu Ala Glu Asp Leu Leu Ser Glu Glu Asp Pro Phe Phe Leu Ala65 70 75 80Glu Leu Leu Tyr Ile Ile Arg Gln Lys Lys Leu Leu Gln His Leu Asn 85 90 95Cys Thr Lys Glu Glu Val Glu Arg Leu Leu Pro Thr Arg Gln Arg Val 100 105 110Ser Leu Phe Arg Asn Leu Leu Tyr Glu Leu Ser Glu Gly Ile Asp Ser 115 120 125Glu Asn Leu Lys Asp Met Ile Phe Leu Leu Lys Asp Ser Leu Pro Lys 130 135 140Thr Glu Met Thr Ser Leu Ser Phe Leu Ala Phe Leu Glu Lys Gln Gly145 150 155 160Lys Ile Asp Glu Asp Asn Leu Thr Cys Leu Glu Asp Leu Cys Lys Thr 165 170 175Val Val Pro Lys Leu Leu Arg Asn Ile Glu Lys Tyr Lys Arg Glu Lys 180 185 190Ala Ile Gln Ile Val Thr Pro Pro Val Asp Lys Glu Ala Glu Ser Tyr 195 200 205Gln Gly Glu Glu Glu Leu Val Ser Gln Thr Asp Val Lys Thr Phe Leu 210 215 220Glu Ala Leu Pro Gln Glu Ser Trp Gln Asn Lys His Ala Gly Ser Asn225 230 235 240Glu Gly Ser Cys Val Gln Asp Glu Ser Glu Pro Gln Arg Pro Leu Cys 245 250 255His Cys Gln Gln Pro Gln Leu Tyr Leu Pro Glu Gly Gln Thr Arg Asn 260 265 270Pro 51522PRTHomo sapiens 51Met Lys Ser Gln Gly Gln His Trp Tyr Ser Ser Ser Asp Lys Asn Cys1 5 10 15Lys Val Ser Phe Arg Glu Lys Leu Leu Ile Ile Asp Ser Asn Leu Gly 20 25 30Val Gln Asp Val Glu Asn Leu Lys Phe Leu Cys Ile Gly Leu Val Pro 35 40 45Asn Lys Lys Leu Glu Lys Ser Ser Ser Ala Ser Asp Val Phe Glu His 50 55 60Leu Leu Ala Glu Asp Leu Leu Ser Glu Glu Asp Pro Phe Phe Leu Ala65 70 75 80Glu Leu Leu Tyr Ile Ile Arg Gln Lys Lys Leu Leu Gln His Leu Asn 85 90 95Cys Thr Lys Glu Glu Val Glu Arg Leu Leu Pro Thr Arg Gln Arg Val 100 105 110Ser Leu Phe Arg Asn Leu Leu Tyr Glu Leu Ser Glu Gly Ile Asp Ser 115 120 125Glu Asn Leu Lys Asp Met Ile Phe Leu Leu Lys Asp Ser Leu Pro Lys 130 135 140Thr Glu Met Thr Ser Leu Ser Phe Leu Ala Phe Leu Glu Lys Gln Gly145 150 155 160Lys Ile Asp Glu Asp Asn Leu Thr Cys Leu Glu Asp Leu Cys Lys Thr 165 170 175Val Val Pro Lys Leu Leu Arg Asn Ile Glu Lys Tyr Lys Arg Glu Lys 180 185 190Ala Ile Gln Ile Val Thr Pro Pro Val Asp Lys Glu Ala Glu Ser Tyr 195 200 205Gln Gly Glu Glu Glu Leu Val Ser Gln Thr Asp Val Lys Thr Phe Leu 210 215 220Glu Ala Leu Pro Gln Glu Ser Trp Gln Asn Lys His Ala Gly Ser Asn225 230 235 240Gly Asn Arg Ala Thr Asn Gly Ala Pro Ser Leu Val Ser Arg Gly Met 245 250 255Gln Gly Ala Ser Ala Asn Thr Leu Asn Ser Glu Thr Ser Thr Lys Arg 260 265 270Ala Ala Val Tyr Arg Met Asn Arg Asn His Arg Gly Leu Cys Val Ile 275 280 285Val Asn Asn His Ser Phe Thr Ser Leu Lys Asp Arg Gln Gly Thr His 290 295 300Lys Asp Ala Glu Ile Leu Ser His Val Phe Gln Trp Leu Gly Phe Thr305 310 315 320Val His Ile His Asn Asn Val Thr Lys Val Glu Met Glu Met Val Leu 325 330 335Gln Lys Gln Lys Cys Asn Pro Ala His Ala Asp Gly Asp Cys Phe Val 340 345 350Phe Cys Ile Leu Thr His Gly Arg Phe Gly Ala Val Tyr Ser Ser Asp 355 360 365Glu Ala Leu Ile Pro Ile Arg Glu Ile Met Ser His Phe Thr Ala Leu 370 375 380Gln Cys Pro Arg Leu Ala Glu Lys Pro Lys Leu Phe Phe Ile Gln Ala385 390 395 400Cys Gln Gly Glu Glu Ile Gln Pro Ser Val Ser Ile Glu Ala Asp Ala 405 410 415Leu Asn Pro Glu Gln Ala Pro Thr Ser Leu Gln Asp Ser Ile Pro Ala 420 425 430Glu Ala Asp Phe Leu Leu Gly Leu Ala Thr Val Pro Gly Tyr Val Ser 435 440 445Phe Arg His Val Glu Glu Gly Ser Trp Tyr Ile Gln Ser Leu Cys Asn 450 455 460His Leu Lys Lys Leu Val Pro Arg His Glu Asp Ile Leu Ser Ile Leu465 470 475 480Thr Ala Val Asn Asp Asp Val Ser Arg Arg Val Asp Lys Gln Gly Thr 485 490 495Lys Lys Gln Met Pro Gln Pro Ala Phe Thr Leu Arg Lys Lys Leu Val 500 505 510Phe Pro Val Pro Leu Asp Ala Leu Ser Leu 515 5205219DNAArtificial SequenceTarget sequence 52tatcctgatg ttgcttggc 195319DNAArtificial SequenceTarget sequence 53gtggcgcatg atgtccctc 195419DNAArtificial SequenceTarget sequence 54tcaagggcca gtatcaagc 195519DNAArtificial SequenceTarget sequence 55gatgtcacca atcctttgc 195619DNAArtificial SequenceTarget sequence 56tcatgtggat cctctcagc 195719DNAArtificial SequenceTarget sequence 57cttcatcaag gtgttgggc 195819DNAArtificial SequenceTarget sequence 58gccttgtcat ttgacaacc 195919DNAArtificial SequenceTarget sequence 59ctacaaggtc cctaccttc 196019DNAArtificial SequenceTarget sequence 60ggtcatgttg gcagaactc 196119DNAArtificial SequenceTarget sequence 61gaagatcaag ccacccttc 196219DNAArtificial SequenceTarget sequence 62ggcattgaca atcggactc 196319DNAArtificial SequenceTarget sequence 63ttcctacttg accgagctc 196419DNAArtificial SequenceTarget sequence 64acttcctact tgaccgagc 196519DNAArtificial SequenceTarget sequence 65attaaagcgg ccaacgtcc 196619DNAArtificial SequenceTarget sequence 66ctggcccatt agatgaaac 196719DNAArtificial SequenceTarget sequence 67agccatcaag gaaacttcc 196819DNAArtificial SequenceTarget sequence 68gctagaaatg aggtacgtc 196919DNAArtificial SequenceTarget sequence 69gtgtttgcta acagtgtcc 197019DNAArtificial SequenceTarget sequence 70gccagccaag taacaaagc 197119DNAArtificial SequenceTarget sequence 71caaagcagat agctttggc 197219DNAArtificial SequenceTarget sequence 72ctggcagaaa ttgtgaagc 197319DNAArtificial SequenceTarget sequence 73accacgtccc acgaatccc 197419DNAArtificial SequenceTarget sequence 74ggacagtgtc tatatccac 197519DNAArtificial SequenceTarget sequence 75acaccacagc cggactatc 197619DNAArtificial SequenceTarget sequence 76ctcctaagag atactctgc 197719DNAArtificial SequenceTarget sequence 77agttcaaggt gccgtgagc 197819DNAArtificial SequenceTarget sequence 78gttcaaggtg ccgtgagcc 197919DNAArtificial SequenceTarget sequence 79tggcaaagag tatgccgtc 198019DNAArtificial SequenceTarget sequence 80attgcaagga ggttccatc 198119DNAArtificial SequenceTarget sequence 81ggaggttcca tcttagccc 198219DNAArtificial SequenceTarget sequence 82atcagtctgg tcctagaac 198319DNAArtificial SequenceTarget sequence 83ctcgtcacag gaggagaac 198419DNAArtificial SequenceTarget sequence 84atcaagtgct catgaagac 198519DNAArtificial SequenceTarget sequence 85gaggacatga aagctattc 198619DNAArtificial SequenceTarget sequence 86atccattgtg tacagatgc 198719DNAArtificial SequenceTarget sequence 87ccacatcact gcgatttcc 198819DNAArtificial SequenceTarget sequence 88catcatccca gtcattggc 198919DNAArtificial SequenceTarget sequence 89caacctctat gaggatcgc 199019DNAArtificial SequenceTarget sequence 90tgctctgact gctggcttc 199119DNAArtificial SequenceTarget sequence 91gagatgctgg agaagatgc 199219DNAArtificial SequenceTarget sequence 92atggatgctg tgtgcaagc 199319DNAArtificial SequenceTarget sequence 93ctgcatgtgt acgagattc 199419DNAArtificial SequenceTarget sequence 94gagattccgg ttgagtacc 199519DNAArtificial SequenceTarget sequence 95cagtccagag actcatccc 199619DNAArtificial SequenceTarget sequence 96actggagaaa tgtggacac 199719DNAArtificial SequenceTarget sequence 97gctgagcaac atggagaac 199819DNAArtificial SequenceTarget sequence 98cgcactagga cttatgctc 199919DNAArtificial SequenceTarget sequence 99ttccaagctg gccacattc 1910019DNAArtificial SequenceTarget sequence 100aacctgcctg cgatacctc 1910119DNAArtificial SequenceTarget sequence 101taggacttat gctcaaagc 1910219DNAArtificial SequenceTarget sequence 102cggatagagg aacgtagcc 1910319DNAArtificial SequenceTarget sequence 103tgggttggga agataccgc 1910419DNAArtificial SequenceTarget sequence 104gacatggcag tgattgctc 1910519DNAArtificial SequenceTarget sequence 105aagacatggc agtgattgc 1910619DNAArtificial SequenceTarget sequence 106gtgctggatt ctgccatgc 1910719DNAArtificial SequenceTarget sequence 107ctggcttgct attagtaac 1910819DNAArtificial SequenceTarget sequence 108atcttccgca tctgccaac 1910919DNAArtificial SequenceTarget sequence 109gaggactaaa gcgcctctc 1911019DNAArtificial SequenceTarget sequence 110tgtgcacctt tgttgaacc 1911119DNAArtificial SequenceTarget sequence 111tacagccaac gacccttac 1911219DNAArtificial SequenceTarget sequence 112gccggaagtc ctgtatacc 1911319DNAArtificial SequenceTarget sequence 113agccggaagt cctgtatac 1911419DNAArtificial SequenceTarget sequence 114gatggctcat cacacactc 1911519DNAArtificial SequenceTarget sequence 115cggctcaaga aactggagc 1911619DNAArtificial SequenceTarget sequence 116gttgtctcaa gtgccaggc 1911719DNAArtificial SequenceTarget sequence 117acacgtgctt cctgaatgc 1911819DNAArtificial SequenceTarget sequence 118cctgggaaac acgtgcttc 1911919DNAArtificial SequenceTarget sequence 119cgaccacaac cacatgtgc 1912019DNAArtificial SequenceTarget sequence 120cgtgaccaag atgcccaac 1912119DNAArtificial SequenceTarget sequence 121tgcagctgac aactttcac 1912219DNAArtificial SequenceTarget sequence 122actctgtggg aaccaattc 1912319DNAArtificial SequenceTarget sequence 123attcccacca cagaacgac 1912419DNAArtificial SequenceTarget sequence 124agctaccaga gcgtcttcc 1912519DNAArtificial SequenceTarget sequence 125tctctccatc tgtctctac 1912619DNAArtificial SequenceTarget sequence 126tgatgaactt gtacttcac 1912719DNAArtificial SequenceTarget sequence 127cgtgatactg ctttacacc 1912819DNAArtificial SequenceTarget sequence 128cattaatgac acgctcttc 1912919DNAArtificial SequenceTarget sequence 129atcccactga tgacaccac 1913019DNAArtificial SequenceTarget sequence 130atcacaggca tcatcatcc 1913119DNAArtificial SequenceTarget sequence 131aagagcacca aagcagctc 1913219DNAArtificial SequenceTarget sequence 132gtccagttct tcagcagac 1913319DNAArtificial SequenceTarget sequence 133cttcttctat gatgaacac 1913419DNAArtificial SequenceTarget sequence 134aagtcagacc tcaaacagc 1913519DNAArtificial SequenceTarget sequence 135caccgattcc cagaagatc 1913619DNAArtificial SequenceTarget sequence 136atctgctgca taccgctac 1913719DNAArtificial SequenceTarget sequence 137catgcagttc atcagccac
1913819DNAArtificial SequenceTarget sequence 138tgcagtgctg tatcaaagc 1913919DNAArtificial SequenceTarget sequence 139ctcctgacct caagtgatc 1914019DNAArtificial SequenceTarget sequence 140tctgcatagg attggtccc 1914119DNAArtificial SequenceTarget sequence 141catctcttgg cagaggatc 1914219DNAArtificial SequenceTarget sequence 142gagtggacaa acagggaac 1914319DNAArtificial SequenceTarget sequence 143tgcttcgtgt tctgtattc 1914419DNAArtificial SequenceTarget sequence 144acagatctca gcaacctcc 1914519DNAArtificial SequenceTarget sequence 145gacggaaacc tccctttac 1914619DNAArtificial SequenceTarget sequence 146aaccacagct ttacctccc 1914712DNAArtificial SequenceTarget sequence 147gtttgctata ac 1214819DNAArtificial SequenceTarget sequence 148ttcaatgccc ggcgtaagc 1914919DNAArtificial SequenceTarget sequence 149tctaaactct gaaaccagc 1915019DNAArtificial SequenceTarget sequence 150atggtggttc agggaattc 1915119DNAArtificial SequenceTarget sequence 151aggtgagtcc atcagaaac 1915219DNAArtificial SequenceTarget sequence 152agggtcttgc tctgtcacc 1915319DNAArtificial SequenceTarget sequence 153ggatgggaca aagacagac 1915419DNAArtificial SequenceTarget sequence 154gaggcagcag aacctgttc 1915519DNAArtificial SequenceTarget sequence 155ctttgttgaa ccatcacac 1915619DNAArtificial SequenceTarget sequence 156ggcatcagaa ggagaaatc 1915719DNAArtificial SequenceTarget sequence 157atgatgtgtg ccacacacc 1915819DNAArtificial SequenceTarget sequence 158cggcacttta cagagaagc 1915919DNAArtificial SequenceTarget sequence 159gagctgtgac ttgtggtcc 1916019DNAArtificial SequenceTarget sequence 160ctccgagatg tgattgctc 1916119DNAArtificial SequenceTarget sequence 161cattgcgcac agagacctc 1916219DNAArtificial SequenceTarget sequence 162ctgaacaact cctgtaccc 1916319DNAArtificial SequenceTarget sequence 163gacctcatct ccaagctcc 1916419DNAArtificial SequenceTarget sequence 164ggagccgagc tttagaccc 1916519DNAArtificial SequenceTarget sequence 165ctactacacc tctttgtac 1916619DNAArtificial SequenceTarget sequence 166atgtggacac aacaagggc 1916719DNAArtificial SequenceTarget sequence 167tgtggacgac tcgcgcatc 1916819DNAArtificial SequenceTarget sequence 168cgagcacgta tgatctctc 1916919DNAArtificial SequenceTarget sequence 169agggctgcat tctgtgatc 1917019DNAArtificial SequenceTarget sequence 170ctgcacaata tgactcctc 1917119DNAArtificial SequenceTarget sequence 171cctcatcaac agcctcttc 1917219DNAArtificial SequenceTarget sequence 172atcggtgtca ccgtgatac 1917319DNAArtificial SequenceTarget sequence 173ccaggagcct cttccatac 1917419DNAArtificial SequenceTarget sequence 174tcgattccga gctgtcttc 1917519DNAArtificial SequenceTarget sequence 175gacagcaaga ttgtggacc 1917650RNAArtificial SequencesiRNA 176gccaagcaac aucaggauag uuugcuauaa cuauccugau guugcuuggc 5017719RNAArtificial SequencesiRNA 177gccaagcaac aucaggaua 1917819RNAArtificial SequencesiRNA 178uauccugaug uugcuuggc 1917919DNAArtificial SequenceTarget sequence 179gctgaccctg aagttcatc 1918019DNAArtificial SequenceTarget sequence 180gctgaccctg aagttcatc 1918127DNAArtificial SequencePCR primers 181gcgaagcttg cggcatggac gaactgt 2718227DNAArtificial SequencePCR primers 182gcgggatccc aggcgtcacc cccttag 2718325DNAArtificial SequencePCR primers 183gttctggggt gtggtgtctc acagc 2518425DNAArtificial SequencePCR primers 184caaactgagc cacatcaggc actcc 2518526DNAArtificial SequencePCR primers 185ccagtctgaa agtgactgct gtgtgg 2618626DNAArtificial SequencePCR primers 186caactggaca tttgtgacct gcatcc 2618719DNAArtificial SequenceTarget sequence 187gctgaccctg aagttcatc 1918820DNAArtificial SequencePCR primers 188ggtgggaggt ctatataagc 2018922DNAArtificial SequencePCR primers 189ggacaaacca caactagaat gc 2219018DNAArtificial SequencePCR primers 190ccccaggcac tggtgttg 1819119DNAArtificial SequencePCR primers 191acggaccact tggccttct 1919225DNAArtificial SequencePCR primers 192ccggtttttc aaagggaata agtac 2519324DNAArtificial SequencePCR primers 193ttcacagttc tagggaagcc aaag 2419418DNAArtificial SequencePCR primers 194cagcatccgt gggtcaca 1819520DNAArtificial SequencePCR primers 195ttcaccgctg ccttaagctt 2019624DNAArtificial SequencePCR primers 196tgaggacgac ctatttgagt ccat 2419722DNAArtificial SequencePCR primers 197gggattcttc gtcatgaaag ct 2219823DNAArtificial SequencePCR primers 198ctgcgaagct gtgaatccta ctc 2319920DNAArtificial SequencePCR primers 199ggcatcctgc tggctgtatc 2020027DNAArtificial SequencePCR primers 200tcctggcaga actcctctat atcatac 2720125DNAArtificial SequencePCR primers 201tgacagttcg tagagcaggt ttcta 2520222DNAArtificial SequencePCR primers 202gaacttgtac ttcacgcagg tg 2220322DNAArtificial SequencePCR primers 203caacaggaaa acaaggctga tg 2220422DNAArtificial SequencePCR primers 204tgcgtggtcc cttctttatc ac 2220520DNAArtificial SequencePCR primers 205gccatggaga cgctcttcag 2020628DNAArtificial SequencePCR primers 206cattaaatga actcctacat aggaaaac 2820720DNAArtificial SequencePCR primers 207agggcaattt catgcaggat 2020823DNAArtificial SequencePCR primers 208ggagtgtctc tgtcaaaagt gga 2320926DNAArtificial SequencePCR primers 209acccattttg atagagctcc tacatt 2621020DNAArtificial SequencePCR primers 210gacattaaag cggccaacgt 2021118DNAArtificial SequencePCR primers 211ctcgggtgcc atccagaa 1821221DNAArtificial SequencePCR primers 212gcgagaaaga cgaagagctg c 2121318DNAArtificial SequencePCR primers 213gcctggctct gctgcatt 1821425DNAArtificial SequencePCR primers 214cagtgaaatt tatccacaat cacac 2521522DNAArtificial SequencePCR primers 215agagctgagg ccagtccaat at 2221622DNAArtificial SequencePCR primers 216ggtgtaggga agagtgccat ga 2221721DNAArtificial SequencePCR primers 217gcatcttcaa tggtgggatc a 2121820DNAArtificial SequencePCR primers 218tggtcgcctg tgtcttctca 2021925DNAArtificial SequencePCR primers 219gtggtatctt tcaacgtctg tcctc 2522020DNAArtificial SequencePCR primers 220gaggaaggcg gtggtagtga 2022122DNAArtificial SequencePCR primers 221ctcaaccgga atctcgtaca ca 2222224DNAArtificial SequencePCR primers 222tgggagataa cttgggtctc tgat 2422321DNAArtificial SequencePCR primers 223aagccaattc tggtcgagct t 2122421DNAArtificial SequencePCR primers 224agggagccta tgccaaagtt c 2122522DNAArtificial SequencePCR primers 225ctcgatgatt ttgacggcat ac 2222623DNAArtificial SequencePCR primers 226gaggaagctc ctgaaggtca aac 2322720DNAArtificial SequencePCR primers 227caaccactgc cttgtccatc 2022821DNAArtificial SequencePCR primers 228agccagctgc agttcttcct t 2122923DNAArtificial SequencePCR primers 229tcacagcgtc tcttgactga aag 2323020DNAArtificial SequencePCR primers 230ggtgggaggt ctatataagc 2023122DNAArtificial SequencePCR primers 231ggacaaacca caactagaat gc 2223231PRTArtificial SequenceGPCR Domain 232Met Asn Ser Thr Leu Asp Gly Asn Gln Ser Ser His Pro Phe Cys Leu1 5 10 15Leu Ala Phe Gly Tyr Leu Glu Thr Val Asn Phe Cys Leu Leu Glu 20 25 3023323PRTArtificial SequenceGPCR Domain 233Val Leu Ile Ile Val Phe Leu Thr Val Leu Ile Ile Ser Gly Asn Ile1 5 10 15Ile Val Ile Phe Val Phe His 2023411PRTArtificial SequenceGPCR Domain 234Cys Ala Pro Leu Leu Asn His His Thr Thr Ser1 5 1023523PRTArtificial SequenceGPCR Domain 235Tyr Phe Ile Gln Thr Met Ala Tyr Ala Asp Leu Phe Val Gly Val Ser1 5 10 15Cys Val Val Pro Ser Leu Ser 2023614PRTArtificial SequenceGPCR Domain 236Leu Leu His His Pro Leu Pro Val Glu Glu Ser Leu Thr Cys1 5 1023723PRTArtificial SequenceGPCR Domain 237Gln Ile Phe Gly Phe Val Val Ser Val Leu Lys Ser Val Ser Met Ala1 5 10 15Ser Leu Ala Cys Ile Ser Ile 2023820PRTArtificial SequenceGPCR Domain 238Asp Arg Tyr Ile Ala Ile Thr Lys Pro Leu Thr Tyr Asn Thr Leu Val1 5 10 15Thr Pro Trp Arg 2023923PRTArtificial SequenceGPCR Domain 239Leu Arg Leu Cys Ile Phe Leu Ile Trp Leu Tyr Ser Thr Leu Val Phe1 5 10 15Leu Pro Ser Phe Phe His Trp 2024024PRTArtificial SequenceGPCR Domain 240Gly Lys Pro Gly Tyr His Gly Asp Val Phe Gln Trp Cys Ala Glu Ser1 5 10 15Trp His Thr Asp Ser Tyr Phe Thr 2024123PRTArtificial SequenceGPCR Domain 241Leu Phe Ile Val Met Met Leu Tyr Ala Pro Ala Ala Leu Ile Val Cys1 5 10 15Phe Thr Tyr Phe Asn Ile Phe 2024236PRTArtificial SequenceGPCR Domain 242Arg Ile Cys Gln Gln His Thr Lys Asp Ile Ser Glu Arg Gln Ala Arg1 5 10 15Phe Ser Ser Gln Ser Gly Glu Thr Gly Glu Val Gln Ala Cys Pro Asp 20 25 30Lys Arg Tyr Ala 3524323PRTArtificial SequenceGPCR Domain 243Met Val Leu Phe Arg Ile Thr Ser Val Phe Tyr Ile Leu Trp Leu Pro1 5 10 15Tyr Ile Ile Tyr Phe Leu Leu 202449PRTArtificial SequenceGPCR Domain 244Glu Ser Ser Thr Gly His Ser Asn Arg1 524523PRTArtificial SequenceGPCR Domain 245Phe Ala Ser Phe Leu Thr Thr Trp Leu Ala Ile Ser Asn Ser Phe Cys1 5 10 15Asn Cys Val Ile Tyr Ser Leu 2024643PRTArtificial SequenceGPCR Domain 246Ser Asn Ser Val Phe Gln Arg Gly Leu Lys Arg Leu Ser Gly Ala Met1 5 10 15Cys Thr Ser Cys Ala Ser Gln Thr Thr Ala Asn Asp Pro Tyr Thr Val 20 25 30Arg Ser Lys Gly Pro Leu Asn Gly Cys His Ile 35 402479PRTArtificial SequenceGPCR Domain 247Met Ala Trp Arg Gly Ala Gly Pro Ser1 524823PRTArtificial SequenceGPCR Domain 248Val Pro Gly Ala Pro Gly Gly Val Gly Leu Ser Leu Gly Leu Leu Leu1 5 10 15Gln Leu Leu Leu Leu Leu Gly 20249188PRTArtificial SequenceGPCR Domain 249Pro Ala Arg Gly Phe Gly Asp Glu Glu Glu Arg Arg Cys Asp Pro Ile1 5 10 15Arg Ile Ser Met Cys Gln Asn Leu Gly Tyr Asn Val Thr Lys Met Pro 20 25 30Asn Leu Val Gly His Glu Leu Gln Thr Asp Ala Glu Leu Gln Leu Thr 35 40 45Thr Phe Thr Pro Leu Ile Gln Tyr Gly Cys Ser Ser Gln Leu Gln Phe 50 55 60Phe Leu Cys Ser Val Tyr Val Pro Met Cys Thr Glu Lys Ile Asn Ile65 70 75 80Pro Ile Gly Pro Cys Gly Gly Met Cys Leu Ser Val Lys Arg Arg Cys 85 90 95Glu Pro Val Leu Lys Glu Phe Gly Phe Ala Trp Pro Glu Ser Leu Asn 100 105 110Cys Ser Lys Phe Pro Pro Gln Asn Asp His Asn His Met Cys Met Glu 115 120 125Gly Pro Gly Asp Glu Glu Val Pro Leu Pro His Lys Thr Pro Ile Gln 130 135 140Pro Gly Glu Glu Cys His Ser Val Gly Thr Asn Ser Asp Gln Tyr Ile145 150 155 160Trp Val Lys Arg Ser Leu Asn Cys Val Leu Lys Cys Gly Tyr Asp Ala 165 170 175Gly Leu Tyr Ser Arg Ser Ala Lys Glu Phe Thr Asp 180 18525023PRTArtificial SequenceGPCR Domain 250Ile Trp Met Ala Val Trp Ala Ser Leu Cys Phe Ile Ser Thr Ala Phe1 5 10 15Thr Val Leu Thr Phe Leu Ile 202519PRTArtificial SequenceGPCR Domain 251Asp Ser Ser Arg Phe Ser Tyr Pro Glu1 525223PRTArtificial SequenceGPCR Domain 252Arg Pro Ile Ile Phe Leu Ser Met Cys Tyr Asn Ile Tyr Ser Ile Ala1 5 10 15Tyr Ile Val Arg Leu Thr Val 2025325PRTArtificial SequenceGPCR Domain 253Gly Arg Glu Arg Ile Ser Cys Asp Phe Glu Glu Ala Ala Glu Pro Val1 5 10 15Leu Ile Gln Glu Gly Leu Lys Asn Thr 20 2525423PRTArtificial SequenceGPCR Domain 254Gly Cys Ala Ile Ile Phe Leu Leu Met Tyr Phe Phe Gly Met Ala Ser1 5 10 15Ser Ile Trp Trp Val Ile Leu 2025570PRTArtificial SequenceGPCR Domain 255Thr Leu Thr Trp Phe Leu Ala Ala Gly Leu Lys Trp Gly His Glu Ala1 5 10 15Ile Glu Met His Ser Ser Tyr Phe His Ile Ala Ala Trp Ala Ile Pro 20 25 30Ala Val Lys Thr Ile Val Ile Leu Ile Met Arg Leu Val Asp Ala Asp 35 40 45Glu Leu Thr Gly Leu Cys Tyr Val Gly Asn Gln Asn Leu Asp Ala Leu 50 55 60Thr Gly Phe Val Val Ala65 7025623PRTArtificial SequenceGPCR Domain 256Pro Leu Phe Thr Tyr Leu Val Ile Gly Thr Leu Phe Ile Ala Ala Gly1 5 10 15Leu Val Ala Leu Phe Lys Ile 2025720PRTArtificial SequenceGPCR Domain 257Arg Ser Asn Leu Gln Lys Asp Gly Thr Lys Thr Asp Lys Leu Glu Arg1 5 10 15Leu Met Val Lys 2025823PRTArtificial SequenceGPCR Domain 258Ile Gly Val Phe Ser Val Leu Tyr Thr Val Pro Ala Thr Cys Val Ile1 5 10 15Ala Cys Tyr Phe Tyr Glu Ile
2025914PRTArtificial SequenceGPCR Domain 259Ser Asn Trp Ala Leu Phe Arg Tyr Ser Ala Asp Asp Ser Asn1 5 1026023PRTArtificial SequenceGPCR Domain 260Met Ala Val Glu Met Leu Lys Ile Phe Met Ser Leu Leu Val Gly Ile1 5 10 15Thr Ser Gly Met Trp Ile Trp 2026141PRTArtificial SequenceGPCR Domain 261Ser Ala Lys Thr Leu His Thr Trp Gln Lys Cys Ser Asn Arg Leu Val1 5 10 15Asn Ser Gly Lys Val Lys Arg Glu Lys Arg Gly Asn Gly Trp Val Lys 20 25 30Pro Gly Lys Gly Ser Glu Thr Val Val 35 4026245PRTArtificial SequenceGPCR Domain 262Met Arg Pro Glu Arg Pro Arg Pro Arg Gly Ser Ala Pro Gly Pro Met1 5 10 15Glu Thr Pro Pro Trp Asp Pro Ala Arg Asn Asp Ser Leu Pro Pro Thr 20 25 30Leu Thr Pro Ala Val Pro Pro Tyr Val Lys Leu Gly Leu 35 40 4526323PRTArtificial SequenceGPCR Domain 263Thr Val Val Tyr Thr Val Phe Tyr Ala Leu Leu Phe Val Phe Ile Tyr1 5 10 15Val Gln Leu Trp Leu Val Leu 2026412PRTArtificial SequenceGPCR Domain 264Arg Tyr Arg His Lys Arg Leu Ser Tyr Gln Ser Val1 5 1026520PRTArtificial SequenceGPCR Domain 265Phe Leu Phe Leu Cys Leu Phe Trp Ala Ser Leu Arg Thr Val Leu Phe1 5 10 15Ser Phe Tyr Phe 2026614PRTArtificial SequenceGPCR Domain 266Lys Asp Phe Val Ala Ala Asn Ser Leu Ser Pro Phe Val Phe1 5 1026723PRTArtificial SequenceGPCR Domain 267Trp Leu Leu Tyr Cys Phe Pro Val Cys Leu Gln Phe Phe Thr Leu Thr1 5 10 15Leu Met Asn Leu Tyr Phe Thr 2026820PRTArtificial SequenceGPCR Domain 268Gln Val Ile Phe Lys Ala Lys Ser Lys Tyr Ser Pro Glu Leu Leu Lys1 5 10 15Tyr Arg Leu Pro 2026923PRTArtificial SequenceGPCR Domain 269Leu Tyr Leu Ala Ser Leu Phe Ile Ser Leu Val Phe Leu Leu Val Asn1 5 10 15Leu Thr Cys Ala Val Leu Val 2027014PRTArtificial SequenceGPCR Domain 270Lys Thr Gly Asn Trp Glu Arg Lys Val Ile Val Ser Val Arg1 5 1027123PRTArtificial SequenceGPCR Domain 271Val Ala Ile Asn Asp Thr Leu Phe Val Leu Cys Ala Val Ser Leu Ser1 5 10 15Ile Cys Leu Tyr Lys Ile Ser 2027220PRTArtificial SequenceGPCR Domain 272Lys Met Ser Leu Ala Asn Ile Tyr Leu Glu Ser Lys Gly Ser Ser Val1 5 10 15Cys Gln Val Thr 2027323PRTArtificial SequenceGPCR Domain 273Ala Ile Gly Val Thr Val Ile Leu Leu Tyr Thr Ser Arg Ala Cys Tyr1 5 10 15Asn Leu Phe Ile Leu Ser Phe 2027430PRTArtificial SequenceGPCR Domain 274Ser Gln Asn Lys Ser Val His Ser Phe Asp Tyr Asp Trp Tyr Asn Val1 5 10 15Ser Asp Gln Ala Asp Leu Lys Asn Gln Leu Gly Asp Ala Gly 20 25 3027523PRTArtificial SequenceGPCR Domain 275Tyr Val Leu Phe Gly Val Val Leu Phe Val Trp Glu Leu Leu Pro Thr1 5 10 15Thr Leu Val Val Tyr Phe Phe 2027686PRTArtificial SequenceGPCR Domain 276Arg Val Arg Asn Pro Thr Lys Asp Leu Thr Asn Pro Gly Met Val Pro1 5 10 15Ser His Gly Phe Ser Pro Arg Ser Tyr Phe Phe Asp Asn Pro Arg Arg 20 25 30Tyr Asp Ser Asp Asp Asp Leu Ala Trp Asn Ile Ala Pro Gln Gly Leu 35 40 45Gln Gly Gly Phe Ala Pro Asp Tyr Tyr Asp Trp Gly Gln Gln Thr Asn 50 55 60Ser Phe Leu Ala Gln Ala Gly Thr Leu Gln Asp Ser Thr Leu Asp Pro65 70 75 80Asp Lys Pro Ser Leu Gly 8527795PRTArtificial SequenceGPCR Domain 277Met Ala Glu Ala Ile Thr Tyr Ala Asp Leu Arg Phe Val Lys Ala Pro1 5 10 15Leu Lys Lys Ser Ile Ser Ser Arg Leu Gly Gln Asp Pro Gly Ala Asp 20 25 30Asp Asp Gly Glu Ile Thr Tyr Glu Asn Val Gln Val Pro Ala Val Leu 35 40 45Gly Val Pro Ser Ser Leu Ala Ser Ser Val Leu Gly Asp Lys Ala Ala 50 55 60Val Lys Ser Glu Gln Pro Thr Ala Ser Trp Arg Ala Val Thr Ser Pro65 70 75 80Ala Val Gly Arg Ile Leu Pro Cys Arg Thr Thr Cys Leu Arg Tyr 85 90 9527823PRTArtificial SequenceGPCR Domain 278Leu Leu Leu Gly Leu Leu Leu Thr Cys Leu Leu Leu Gly Val Thr Ala1 5 10 15Ile Cys Leu Gly Val Arg Tyr 20279241PRTArtificial SequenceGPCR Domain 279Leu Gln Val Ser Gln Gln Leu Gln Gln Thr Asn Arg Val Leu Glu Val1 5 10 15Thr Asn Ser Ser Leu Arg Gln Gln Leu Arg Leu Lys Ile Thr Gln Leu 20 25 30Gly Gln Ser Ala Glu Asp Leu Gln Gly Ser Arg Arg Glu Leu Ala Gln 35 40 45Ser Gln Glu Ala Leu Gln Val Glu Gln Arg Ala His Gln Ala Ala Glu 50 55 60Gly Gln Leu Gln Ala Cys Gln Ala Asp Arg Gln Lys Thr Lys Glu Thr65 70 75 80Leu Gln Ser Glu Glu Gln Gln Arg Arg Ala Leu Glu Gln Lys Leu Ser 85 90 95Asn Met Glu Asn Arg Leu Lys Pro Phe Phe Thr Cys Gly Ser Ala Asp 100 105 110Thr Cys Cys Pro Ser Gly Trp Ile Met His Gln Lys Ser Cys Phe Tyr 115 120 125Ile Ser Leu Thr Ser Lys Asn Trp Gln Glu Ser Gln Lys Gln Cys Glu 130 135 140Thr Leu Ser Ser Lys Leu Ala Thr Phe Ser Glu Ile Tyr Pro Gln Ser145 150 155 160His Ser Tyr Tyr Phe Leu Asn Ser Leu Leu Pro Asn Gly Gly Ser Gly 165 170 175Asn Ser Tyr Trp Thr Gly Leu Ser Ser Asn Lys Asp Trp Lys Leu Thr 180 185 190Asp Asp Thr Gln Arg Thr Arg Thr Tyr Ala Gln Ser Ser Lys Cys Asn 195 200 205Lys Val His Lys Thr Trp Ser Trp Trp Thr Leu Glu Ser Glu Ser Cys 210 215 220Arg Ser Ser Leu Pro Tyr Ile Cys Glu Met Thr Ala Phe Arg Phe Pro225 230 235 240Asp280145PRTArtificial SequenceGPCR Domain 280Met Ser Pro Ser Gly Arg Leu Cys Leu Leu Thr Ile Val Gly Leu Ile1 5 10 15Leu Pro Thr Arg Gly Gln Thr Leu Lys Asp Thr Thr Ser Ser Ser Ser 20 25 30Ala Asp Ser Thr Ile Met Asp Ile Gln Val Pro Thr Arg Ala Pro Asp 35 40 45Ala Val Tyr Thr Glu Leu Gln Pro Thr Ser Pro Thr Pro Thr Trp Pro 50 55 60Ala Asp Glu Thr Pro Gln Pro Gln Thr Gln Thr Gln Gln Leu Glu Gly65 70 75 80Thr Asp Gly Pro Leu Val Thr Asp Pro Glu Thr His Lys Ser Thr Lys 85 90 95Ala Ala His Pro Thr Asp Asp Thr Thr Thr Leu Ser Glu Arg Pro Ser 100 105 110Pro Ser Thr Asp Val Gln Thr Asp Pro Gln Thr Leu Lys Pro Ser Gly 115 120 125Phe His Glu Asp Asp Pro Phe Phe Tyr Asp Glu His Thr Leu Arg Lys 130 135 140Arg14528119PRTArtificial SequenceGPCR Domain 281Gly Leu Leu Val Ala Ala Val Leu Phe Ile Thr Gly Ile Ile Ile Leu1 5 10 15Thr Ser Gly28214PRTArtificial SequenceGPCR Domain 282Lys Cys Arg Gln Leu Ser Arg Leu Cys Arg Asn Arg Cys Arg1 5 10283116PRTArtificial SequenceGPCR Domain 283Met Cys Thr Glu Lys Ile Asn Ile Pro Ile Gly Pro Cys Gly Gly Met1 5 10 15Cys Leu Ser Val Lys Arg Arg Cys Glu Pro Val Leu Lys Glu Phe Gly 20 25 30Phe Ala Trp Pro Glu Ser Leu Asn Cys Ser Lys Phe Pro Pro Gln Asn 35 40 45Asp His Asn His Met Cys Met Glu Gly Pro Gly Asp Glu Glu Val Pro 50 55 60Leu Pro His Lys Thr Pro Ile Gln Pro Gly Glu Glu Cys His Ser Val65 70 75 80Gly Thr Asn Ser Asp Gln Tyr Ile Trp Val Lys Arg Ser Leu Asn Cys 85 90 95Val Leu Lys Cys Gly Tyr Asp Ala Gly Leu Tyr Ser Arg Ser Ala Lys 100 105 110Glu Phe Thr Asp 11528423PRTArtificial SequenceGPCR Domain 284Ile Trp Met Ala Val Trp Ala Ser Leu Cys Phe Ile Ser Thr Ala Phe1 5 10 15Thr Val Leu Thr Phe Leu Ile 202859PRTArtificial SequenceGPCR Domain 285Asp Ser Ser Arg Phe Ser Tyr Pro Glu1 528623PRTArtificial SequenceGPCR Domain 286Arg Pro Ile Ile Phe Leu Ser Met Cys Tyr Asn Ile Tyr Ser Ile Ala1 5 10 15Tyr Ile Val Arg Leu Thr Val 2028725PRTArtificial SequenceGPCR Domain 287Gly Arg Glu Arg Ile Ser Cys Asp Phe Glu Glu Ala Ala Glu Pro Val1 5 10 15Leu Ile Gln Glu Gly Leu Lys Asn Thr 20 2528823PRTArtificial SequenceGPCR Domain 288Gly Cys Ala Ile Ile Phe Leu Leu Met Tyr Phe Phe Gly Met Ala Ser1 5 10 15Ser Ile Trp Trp Val Ile Leu 2028970PRTArtificial SequenceGPCR Domain 289Thr Leu Thr Trp Phe Leu Ala Ala Gly Leu Lys Trp Gly His Glu Ala1 5 10 15Ile Glu Met His Ser Ser Tyr Phe His Ile Ala Ala Trp Ala Ile Pro 20 25 30Ala Val Lys Thr Ile Val Ile Leu Ile Met Arg Leu Val Asp Ala Asp 35 40 45Glu Leu Thr Gly Leu Cys Tyr Val Gly Asn Gln Asn Leu Asp Ala Leu 50 55 60Thr Gly Phe Val Val Ala65 7029023PRTArtificial SequenceGPCR Domain 290Pro Leu Phe Thr Tyr Leu Val Ile Gly Thr Leu Phe Ile Ala Ala Gly1 5 10 15Leu Val Ala Leu Phe Lys Ile 2029120PRTArtificial SequenceGPCR Domain 291Arg Ser Asn Leu Gln Lys Asp Gly Thr Lys Thr Asp Lys Leu Glu Arg1 5 10 15Leu Met Val Lys 2029223PRTArtificial SequenceGPCR Domain 292Ile Gly Val Phe Ser Val Leu Tyr Thr Val Pro Ala Thr Cys Val Ile1 5 10 15Ala Cys Tyr Phe Tyr Glu Ile 2029314PRTArtificial SequenceGPCR Domain 293Ser Asn Trp Ala Leu Phe Arg Tyr Ser Ala Asp Asp Ser Asn1 5 1029423PRTArtificial SequenceGPCR Domain 294Met Ala Val Glu Met Leu Lys Ile Phe Met Ser Leu Leu Val Gly Ile1 5 10 15Thr Ser Gly Met Trp Ile Trp 2029541PRTArtificial SequenceGPCR Domain 295Ser Ala Lys Thr Leu His Thr Trp Gln Lys Cys Ser Asn Arg Leu Val1 5 10 15Asn Ser Gly Lys Val Lys Arg Glu Lys Arg Gly Asn Gly Trp Val Lys 20 25 30Pro Gly Lys Gly Ser Glu Thr Val Val 35 4029619DNAArtificial SequenceTarget sequence 296gctgaccctg aagttcatc 19
Patent applications by Nick Vandeghinste, Duffel BE
Patent applications by Peter Herwig Maria Tomme, Gent BE
Patent applications by Reginald Brys, Korbeek-Dijle BE
Patent applications by GALAPAGOS NV
Patent applications in class By measuring the ability to specifically bind a target molecule (e.g., antibody-antigen binding, receptor-ligand binding, etc.)
Patent applications in all subclasses By measuring the ability to specifically bind a target molecule (e.g., antibody-antigen binding, receptor-ligand binding, etc.)