Abstract:
Disclosed are novel splicing variants of the genes associated with prostate cancer risk and survival, particularly splicing variants of PIK3CD, FGFR3, TSC2, RASGRP2, ITGA4, MET, NF1 and BAK1. The disclosure also relates risk assessment, detection, diagnosis, or prognosis of prostate cancer. More specifically, this disclosure relates to the detection of certain splicing variants of PIK3CD, FGFR3, TSC2, RASGRP2, ITGA4, MET, NF1 and BAK1.

Description:
REFERENCE TO RELATED APPLICATION 
     This application is a National Phase of International Application No. PCT/US2012/056346, filed Sep. 20, 2012, which claims priority to U.S. Provisional Patent Application No. 61/536,957, filed Sep. 20, 2011, which is incorporated herein by reference. 
    
    
     GOVERNMENT RIGHTS 
     This invention was made with government support under R01-CA120316,R01-DK056108, and 5U01-CA-116937 awarded by the NIH. The U.S. Government has certain rights in the invention. 
    
    
     FIELD OF THE INVENTION 
     The present invention relates to novel splicing variants of a number of genes associated with prostate cancer risk and survival, and also the risk assessment, detection, diagnosis, or prognosis of prostate cancer (CaP). More specifically, this invention relates to the detection of certain splicing variants in genes PIK3CD, FGFR3, TSC2, ITGA4, MET, NF1, BAK1, and RASGRP2 to determine the risk, detect, diagnose, or prognosticate prostate cancer, particularly in the African American population. Research for the present invention was supported in part by American Cancer Society grant ACS-IRG-08-091-01. 
     BACKGROUND OF THE INVENTION 
     Prostate cancer (PCa) is the most common form of cancer among males. Overwhelming clinical evidence shows that human prostate cancer has the propensity to metastasize to bone, and the disease appears to progress inevitably from androgen dependent to androgen refractory status, leading to increased patient mortality. This prevalent disease is currently the second leading cause of cancer death among men in the U.S. 
     There are striking population (race) disparities in prostate cancer risk and survival outcome borne out of current health statistics data. This is particularly evident between African Americans (AA) and their Caucasian American (CA) counterparts. Epidemiologic studies have shown that higher mortality and recurrence rates of prostate cancer are still seen in AA men even after adjustment for socioeconomic status, environmental factors and health care access. Thus, it is likely that intrinsic biological differences account for some of the cancer disparities. Identifying these differences has been identified as a high-priority research area by the NIH, NCI and the Center to Reduce Cancer Health Disparities (CRCHD). 
     There are currently very few diagnostics methods available for the diagnosis and prevention of prostate cancer, particularly which can be used as predictor of risk and survival in African American population. Thus, the identification of genetic differences between AA and their CA counterparts, that are responsible for predisposition of prostate cancer would provide for a better understanding of the mechanisms of cancer causation (including ethnic and individual susceptibility), and ultimately lead to ways of prostate cancer prevention. 
     SUMMARY OF THE INVENTION 
     Prostate cancer (PCa) is a disease conferred by multiple gene mutations, numerous alternations in gene expression and aberrant changes in genome composition/architecture. The African American (AA) population exhibits higher incidence and mortality rates compared to Caucasian Americans (CA). The present invention, through systematic mRNA expression profiling, characterizes the global mRNA expression profiles in AA and CA prostate tissue samples. A large number of genes are shown to have differential expression between AA and CA patients. Notably, several genes residing within the 5 oncogenic signaling pathways have been identified as exhibiting differential splicing, which includes but not limited to PIK3CD, FGFR3, TSC2, FGFR2, PDGFRA, ITGA4, MET, EPHA3, NF1, RASGRP2, CTNNB1, TSC2, ATM, CDK4, and RB 1 between AA and CA PCa specimens. Quantitative analysis of the expression profiles of PIK3CD, FGFR3, TSC2, RASGRP2, ITGA4, MET, NF1 and BAK1 in prostate samples confirm differential splicing between the AA and CA patients. With certain splicing variants predominantly exist in AA patients. As a non-limiting example, PIK3CD is expressed predominantly as a long variant in CA patients, whereas the AA patient would have higher portion of a short variant. The alternatively spliced short variant of PIK3CD is found to be a more aggressive form. Increasing the short to long variants ratio in a PCa cell line (MDA PCa 2b) that is representative to the AA PCa PIK3CD expression profile, by knocking down PIK3CD long variant expression increases cell proliferation and cell migration. Selectively knocking down the expression of PIK3CD short variant in the same cell line, decreases the short to long variants ratio, and results in marked decrease of cell proliferation and cell migration. Similarly AA predominant variants of FGFR3, TSC2 and RASGRP2 are also shown to be the more aggressive variant. 
     It is thus discovered by the inventors that alternative splicing variants for genes in the oncogenic signaling pathways, such as PIK3CD, FGFR3, TSC2, FGFR2, PDGFRA, ITGA4, MET, EPHA3, NF1, RASGRP2, CTNNB1, TSC2, ATM, CDK4, and RB1 are strong predictors of prostate cancer risk and survival, particularly in the AA patient population. It is thus an aim of the present invention to predict the risk and survival of a patient, by detecting the presence or absence of AA predominant variants of the genes in the oncogenic signaling pathways, particularly for PIK3CD, FGFR3, TSC2, FGFR2, PDGFRA, ITGA4, MET, EPHA3, NF1, RASGRP2, CTNNB1, TSC2, ATM, CDK4, and RB1, and more particularly for PIK3CD, FGFR3, TSC2, RASGRP2, ITGA4, MET, NF1 and BAK1. It is also an aspect of the present invention to utilize relative proportions of splicing variants of a certain gene as a predictor for PCa risk and survival in a patient. 
     Another aspect of the present invention is directed to isolated polynucleotide sequences of novel splicing variants of PIK3CD, FGFR3, TSC2, RASGRP2, ITGA4, MET, NF1 and BAK1. These novel splicing variants are particularly useful for the detection of the presence or absence of splicing variants in these genes that are in oncogenic signaling pathways. Detection of the presence or absence of splicing variants may be by polymerase chain reaction, by oligonucleotide probes hybridization, particularly high throughput DNA micro array analysis, or high throughput DNA sequencing, or any other means known to one skilled in the art. The isolated novel splicing variants sequences are also useful for targeted silencing of certain splicing variants of these genes. Targeted gene silencing may be by siRNA, miRNA, or other complementary RNA constructs. 
     Additionally, polypeptide products of the novel splicing variants of the present invention may be analyzed for determining the presence or absence of certain splicing variants. Mass spectrometry may be used to identify peptide fragments specific to certain splicing variants. Antibodies specifically recognize specific amino acid sequences of the novel splicing variants may be developed for the detection of the protein products of these splicing variants. The antibodies may be monoclonal antibodies, polyclonal antibodies, Fab, single chain antibody, or other engineered antibody constructs known to one skilled in the art. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is a schematic drawing for prostate biopsy core sampling; 
         FIG. 2  shows differentially expressed exons between AA and CA populations; 
         FIG. 3  shows differential splicing events in AA and CA PCa specimens; 
         FIG. 4  shows quantitative RT-PCR validation of differentially expressed exons in AA and CA specimens; 
         FIG. 5  illustrates alternative splicing events were found in various signaling molecules in the cell survival and proliferation pathways; 
         FIG. 6  shows relative expression levels of PIK3CD, FGFR3, TSC2, ITGA4, MET, NF1, BAK1, and RASGRP2 splicing variants; 
         FIG. 7  shows the effect of PIK3CD splicing variants on cell proliferation and invasion; 
         FIG. 8  shows effect of knockdown RASGRP2 splicing variants on cell proliferation and invasion; 
         FIG. 9  shows effect of knockdown PIK3CD “long” variant on the AKT pathway; and 
         FIG. 10  shows 4 novel PIK3CD variants. 
     
    
    
     DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT 
     Alternative splicing dramatically expands the protein coding repertoire of higher eukaryotes. Current estimates suggest that greater than 60% of all human genes have more than one isoform/splice variant. The expression of specific splice variants is regulated in a developmentally and tissue-specific manner (Black DL: Mechanisms of alternative pre-messenger RNA splicing. Annu Rev Biochem 2003, 72:291-336). Alternatively spliced isoforms from the same gene can produce proteins with drastically different properties. For example, the bcl-x gene utilizes different 5′ splice sites, resulting in proteins that have antagonistic functions. The short form of bcl-x promotes apoptosis, while the long form inhibits cell death (Boise L H, Gonzalez-Garcia M, Postema C E, Ding L, Lindsten T, Turka L A, Mao X, Nunez G, Thompson CB: bcl-x, a bcl-2-related gene that functions as a dominant regulator of apoptotic cell death. Cell 1993, 74:597-608). 
     Characterization of Clinical Specimens 
     Needle biopsy cores were collected by GWU Medical Faculty Associates urologists from right-base, left-base, right-mid, left-mid, right-apex, left-apex, right-transition, and left-transition zones of the prostate gland of individual patients presenting with high serum levels (&gt;7 ng/ml) of prostate specific antigen (PSA). A schematic for 18 core biopsy is shown in  FIG. 1 . Collected cores were immediately examined by a board certified PCa pathologist. PCa cores were determined to have a pathologic tumor stage of 2, and Gleason scores ranging from 6-9. There was no significant difference between the two racial groups (AA versus CA) with respect to age and tumor grade. Paired normal biopsy cores were also available from the same patients for genomic analysis (normal cores typically 1-2 cm away from cancer cores and deemed cancer free by pathologists). Each core contains sufficient RNA material for Affymetrix Human Exon 1.0 ST GeneChip profiling (i.e. 1 μg total RNA). 
     Exon Expression Profiling of AA and CA PCa and Normal Specimens 
     Total RNA was isolated from PCa and paired normal prostate cores. Exon profiling was performed on the Affymetrix Human Exon 1.0 ST GeneChip. The GeneChip represents an optimal platform for both expression profiling and splice variant detection (Kwan T, Benovoy D, Dias C, Gurd S, Provencher C, Beaulieu P, Hudson T J, Sladek R, Majewski J: Genome-wide analysis of transcript isoform variation in humans. Nat Genet 2008, 40:225-231; Network TCGAR: Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature 2008, 455:1061-1068), as exon level annotations are derived from empirically determined, highly curated mRNA sequences and ab-initio computational predictions (see www.affymetrix.com/support/technical/whitepapers.affx). The GeneChip contains approximately 5.4 million 5-1 μm features (probes) grouped into 1.4 million probe sets interrogating over one million exon clusters. A 4-way statistical design (t-test with 10% false discovery rate (FDR) for multiple test correction) was employed to identify differentially expressed exons (corresponding to differentially expressed splice variants) in the following comparisons: AA normal vs. CA normal, AA cancer vs. CA cancer, AA cancer vs. AA normal, and CA cancer vs. CA normal. See  FIG. 1A  for comparison of AA cancer vs. CA cancer at the exon level. 
     The inventor through exon level analysis has identified 861 genes (e.g. PIK3CD, FGFR3, TSC2, RASGRP2, ITGA4, MET, NF1 and BAK1) exhibiting differential splicing patterns between the AA and CA populations. Differentially expressed exons between AA and CA populations are shown in  FIG. 2 .  FIG. 2(A)  shows Principle Components Analysis (PCA) plots and clustering analysis of differentially expressed exons between AA and CA PCa specimens. 20 AA and 15 CA PCa specimens were analyzed for global alternative splicing patterns (i.e. differentially expressed exons) using the Affymetrix human Exon 1.0 ST arrays. These splice variants represent candidate markers mediating PCa disparities. An example of a gene exhibiting population-specific splicing is integrin α4 (ITGA4) which has been postulated to be a metastasis suppressor, since blocking its activity with antisense RNA enhances oral squamous carcinoma cell motility (Zhang Y, Lu H, Dazin P, Kapila Y: Functional differences between integrin alpha4 and integrins alpha5/alphaV in modulating the motility of human oral squamous carcinoma cells in response to the V region and heparin-binding domain of fibronectin. Exp Cell Res 2004, 295:48-58.). 
       FIG. 3  shows relative expression of individual exons of PIK3CD, FGFR3, and TSC2 in AA and CA prostate cancers.  FIG. 3( a )  shows PIK3CD (phosphoinositide-3-kinase, catalytic, delta polypeptide) variants expression,  FIG. 3( b )  shows FGFR3 (fibroblast growth factor receptor 3) variants expression, and  FIG. 3( c )  shows TSC2 (tuberous sclerosis 2). Arrows indicate exons that are missing in the AA variant but present in the CA variant for each gene. Specifically, PIK3CD variants that lack exons 10 and 23, FGFR3 variant lack exon 14, and TSC2 variant lacks exon 19 are more prevalent in AA PCa patients. 
       FIG. 4  shows quantitative RT-PCR validation of differentially expressed exons in AA and CA specimens. AA and CA patient samples are analyzed using quantitative RT-PCR, using primers listed in Table 1. Preferential expression of a particular exon in either AA or CA PCa specimens for the PIK3CD, FGFR3, TSC2, ITGA4, MET, NF1, BAK1, and RASGRP2 genes is seen. E1F1AX and PPA1 served as internal RT-PCR control genes, which are expressed equally in AA and CA PCa specimens. 
     
       
         
               
             
               
               
             
           
               
                 TABLE 1 
               
               
                   
               
               
                 Primers for qRT-PCR validations of splice variants 
               
               
                 (-L and -S forms) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 PIK3CD 
                 Primer-f (SEQ ID No. 2): 
               
               
                   
                 CAAACTGAAGGCCCTGAATGA 
               
               
                   
                 Primer-r (SEQ ID No. 3): 
               
               
                   
                 TCTCGGATCATGATGTTGTCG 
               
               
                   
               
               
                 FGFR3 
                 Primer-f (SEQ ID No. 20): 
               
               
                   
                 ACAACGTGATGAAGATCGCA 
               
               
                   
                 Primer-r (SEQ ID No. 21): 
               
               
                   
                 AGGTCGTGTGTGCAGTTGG 
               
               
                   
               
               
                 TSC2 
                 Primer-f (SEQ ID No. 29): 
               
               
                   
                 TTTGACTTCCTGTTGCTGCT 
               
               
                   
                 Primer-r (SEQ ID No. 30): 
               
               
                   
                 TGAGCACTTTATAGCGCAG 
               
               
                   
               
               
                 RASGRP2 
                 Primer-f (SEQ ID No. 38): 
               
               
                   
                 TCACGGTGTCTCTGGATCAGT 
               
               
                   
                 Primer-r (SEQ ID No. 39): 
               
               
                   
                 CCACCATCTTCTCGATGTGCT 
               
               
                   
               
               
                 ITGA4 
                 Primer-f (SEQ ID No. 53): 
               
               
                   
                 TCTTGCTGTTGGGAGTATGAA 
               
               
                   
                 Primer-r (SEQ ID No. 54): 
               
               
                   
                 TGATACTGAGGTCCTCTTCCG 
               
               
                   
               
               
                 MET 
                 Primer-f (SEQ ID No. 66): 
               
               
                   
                 TGGTGGAAAGAACCTCTCAA 
               
               
                   
                 Primer-r (SEQ ID No. 67): 
               
               
                   
                 ATCTTGGCTCACTGCAACCT 
               
               
                   
               
               
                 NF1 
                 Primer-f (SEQ ID No. 71): 
               
               
                   
                 GCATTTTGGAACTGGGTAGAA 
               
               
                   
                 Primer-r (SEQ ID No. 72): 
               
               
                   
                 AACCACCATGGACTGAACAA 
               
               
                   
               
               
                 BAK1 
                 Primer-f (SEQ ID No. 80): 
               
               
                   
                 CCTGTTTGAGAGTGGCATCAA 
               
               
                   
                 Primer-r (SEQ ID No. 81): 
               
               
                   
                 TTGATGCCACTCTCAAACAGG 
               
               
                   
               
             
          
         
       
     
     Recently, genome sequencing efforts as part of the Cancer Genome Atlas Project has demonstrated that a number of genes (e.g. RAS, PTEN, p53, PI3K, APC, etc.) exhibiting frequent mutational hits in cancers can be found primarily residing in 3-5 major signaling pathways (Network TCGAR: Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature 2008, 455:1061-1068; Parsons D W, Jones S, Zhang X, Lin J C, Leary R J, Angenendt P, Mankoo P, Carter H, Siu IM, Gallia G L, et al: An integrated genomic analysis of human glioblastoma multiforme. Science 2008, 321:1807-1812; Ding L, Getz G, Wheeler D A, Mardis E R, McLellan M D, Cibulskis K, Sougnez C, Greulich H, Muzny D M, Morgan M B, et al: Somatic mutations affect key pathways in lung adenocarcinoma. Nature 2008, 455:1069-1075). Of interest from a cancer disparities perspective is our observation that many of these same genes are prone to population-specific splicing patterns.  FIG. 5  indicates genes marked with (AS) define differential alternative splicing events occurring in AA versus CA PCa. (Copy number amplifications (A) and deletions (D) are also indicated). At least 11 out of 26 genes residing in the 5 oncogenic signaling pathways have been identified by the inventors as exhibiting differential splicing between AA and CA PCa specimens. These genes include FGFR2, PDGFRA, MET, EPHA3, NF1, RASGRP2, CTNNB 1, TSC2, ATM, CDK4, and RB 1. The inventors further show that differential mRNA splicing in racial populations plays an important role in cancer health disparities. 
       FIG. 6  shows quantification of differential splicing in PIK3CD, FGFR3, TSC2, ITGA4, MET, NF1, BAK1, and RASGRP2 in AA and CA PCa patients. For each of these genes, one variant is predominant in AA patients. Also, proportions of variants, such as short and long form of PIK3CD are markedly different between AA and CA patients. AA patients have a higher S/L ration than CA patients. 
     Functional Consequences of Splice Variants in PCa Cell Lines Derived from AA and CA Patients 
     Inventors demonstrate that the splice variant (short form or S variant) for phosphoinositide-3 kinase delta (PIK3CD) found in AA PCa specimens encodes a more aggressive version of the gene (i.e. leading to greater proliferation and invasion of cancer cells) compared to the variant counterpart (long form or L variant) found in CA PCa specimens ( FIG. 7 ). In the CA PCa cell line VCaP, the L form is the only variant found, while very little to no expression of the S variant is seen (and hence the reason we refer to the L variant as the ‘CA isoform’) ( FIG. 7A ). The predominant expression of the L variant and very little to no expression of the S variant in the CA PCa cell line is consistent with the CA patient samples (see PIK3CD in  FIG. 4 ). SiRNA-mediated knockdown of the L variant in VCaP cells leads to a decrease in Matrigel invasion and a decrease in proliferation ( FIG. 7A ). By comparison, the AA PCa cell line MDA PCa 2b expresses both an L and S variant, and knockdown of the L variant leads to an increase in Matrigel invasion and an increase in proliferation ( FIG. 7A ). Since VCaP cells express very little to no S variant, targeted siRNA-mediated knockdown of this variant leads to no change in Matrigel invasion and proliferation ( FIG. 7B ). In contrast, targeted knockdown of the S variant in MDA PCa 2b cells leads to decreased Matrigel invasion and decreased proliferation (since the S variant is found almost exclusively in AA patient samples, it is referred to as the ‘AA variant’) ( FIG. 7B ). These data indicate that the balance of S to L isoforms in MDA PCa 2b cells dictates the oncogenic profile of the AA PCa cell line. Namely, knocking down the L variant in MDA PCa 2b cells increases the S/L ratio, leading to a higher proportion of the aggressive S variant and consequently increased invasiveness and proliferation of the cell line. In contrast, knocking down the S variant in MDA PCa 2b cells decreases the S/L ratio, leading to a higher proportion of the less aggressive L variant and consequently decreased invasiveness and proliferation of the cell line. Analogous findings were obtained in MDA PCa 2b cells when the ratio of the ‘AA variant’ (S or b isoform) was increased over the ‘CA variant’ (L or an isoform) for the FGFR3, TSC2. 
     For RASGRP2, the long variant (with exon 10) is common to both AA and CA patients, whereas the short variant (without exon 10) is unique to AA. Targeted knockdown of the long splicing variant in VCaP cells reduced Matrigel invasion and an increase in proliferation ( FIG. 8 ). In contrast, target knockdown of the RASGRP2 long variant in MDA PCa 2b Cells has the opposite effect. 
     Activation of AKT is known to promote cell growth and mRNA translation ( FIG. 9 a   ). When the expression of PIK3CD “long” variant is knocked down by siRNA targeting of Exon 23 in the VCaP cell line, which only expresses the long variant, there is a decrease of phosphorylation of AKT, compared to nonsense siRNA control, and also decrease of phosphorylation of downstream signaling proteins mTOR and S6 ( FIG. 9 b   ). However, in MDA PCa 2b cells, which express the short variant of PIK3CD, knocking down the long variant of PIK3CD markedly increases AKT phosphorylation, both on Thr308 and Ser473, and increases phosphorylation of mTOR and S6 ( FIG. 9 b   ). In other words, increasing S/L variants proportion in MDA PCa 2b cells activates the AKT pathways. 
     The inventor discovered four novel PIK3CD variants ( FIG. 10 ), where variant 1 lacks exon 10 (SEQ ID No. 7), which can be shown as the deletion of nt2430-2592 compared to full length PIK3CD cDNA sequence (SEQ ID. No. 1), variant 2 lacks exon 23 (SEQ ID No. 11, deletion of nt931-1020), variant 3 lacks both exon 10 and 23 (SEQ ID No. 14, deletion of nt931-1020 and nt2430-2592), and variant 4 contains a deletion from nt1329-2627 (SEQ ID No. 16). The nucleotide sequence of PIK3CD full length cDNA sequence is shown in Table 2. Exon 10 and exon 23 are marked with double underline and wave underline, respectively. cDNA sequence of variants 1-4 (SEQ ID Nos. 7, 11, 14, and 16) are shown in Tables 3-6. Exemplary primers across the junctions of the splicing variants (SEQ ID Nos. 6, 10, and 15) that are useful for detecting the presence of these variants are shown in Table 7. Exemplary siRNAs for selective knockdown of PIK3CD full length (targeting exon 23, SEQ ID Nos. 4 and 5)) and variants (targeting exon junctions (SEQ ID Nos. 8, 9, 12, and 13) and deletion junction (SEQ ID Nos. 17 and 18)) are listed in Table 8. 
     The inventor also discovered a novel splicing variant of FGFR3 (fibroblast growth factor receptor 3), which lacks exon 14 (SEQ ID No. 19, Table 10). The nucleotide sequence of FGFR3 full length cDNA sequence (SEQ ID No. 19) is shown in Table 9. Exon 14 is marked with double underline. Exemplary primer across the junction of splicing variant (SEQ ID No. 26) that is useful for detecting the presence of this variant is shown in Table 11. Exemplary siRNAs for selective knockdown of FGFR3 full length (targeting exon 14, SEQ ID NOs. 22 and 23)) and variant (targeting exon junction (SEQ ID Nos. 26 and 27) are listed in Table 12. 
     The inventor also discovered a novel splicing variant of TSC2 (tuberous sclerosis 2), which lacks exon 19 (SEQ ID No. 34, Table 14). The nucleotide sequence of TSC2 full length cDNA sequence (SEQ ID No. 28) is shown in Table 12. Exon 19 is marked with double underline. Exemplary primer across the junction of splicing variant (SEQ ID No. 33) that is useful for detecting the presence of this variant is shown in Table 15. Exemplary siRNAs for selective knockdown of TSC2 full length (targeting exon 19, SEQ ID NOs. 31 and 32)) and variant (targeting exon junction (SEQ ID Nos. 35 and 36) are listed in Table 16. 
     The inventor also discovered two novel splicing variants of RASGRP2 (RAS guanyl-releasing protein 2), which lacks exon 10 (SEQ ID No. 45, Table 18) or exon 11 (SEQ ID No. 49, Table 19). The nucleotide sequence of RASGRP2 full length cDNA sequence (SEQ ID No. 37) is shown in Table 17. Exon 10 is marked with double underline, and exon 11 is marked with wave underline. Exemplary primers across the junctions of the splicing variants (SEQ ID Nos. 44 and 48) that are useful for detecting the presence of these variants are shown in Table 20. Exemplary siRNAs for selective knockdown RASGRP2 full length (targeting exon 10, SEQ ID NOs. 40 and 41, targeting exon 11, SEQ ID NOs. 42 and 43)) and variants (targeting exon junctions (SEQ ID Nos. 46, 47, 50, and 51)) are listed in Table 21. 
     The inventor also discovered a novel splicing variant of ITGA4 (integrin α4), which lacks exon 23 (SEQ ID No. 58, Table 23). The nucleotide sequence of ITGA4 full length cDNA sequence (SEQ ID No. 52) is shown in Table 22. Exon 23 is marked with double underline. Exemplary primer across the junction of splicing variant (SEQ ID No. 57) that is useful for detecting the presence of this variant is shown in Table 24. Exemplary siRNAs for selective knockdown of ITGA4 full length (targeting exon 23, SEQ ID NOs. 55 and 56)) and variant (targeting exon junction (SEQ ID Nos. 59 and 60)) are listed in Table 25. 
     The inventor also discovered a novel splicing variant of MET (MNNG HOS Transforming gene), which include the insertion of non-coding exon 27 (SEQ ID No. 65, Table 27). The nucleotide sequence of MET full length cDNA sequence (SEQ ID No. 62) is shown in Table 26. Exon 27 is marked with double underline. Exemplary primer across junctions of full length variant (SEQ ID No. 61) is shown in Table 28. Exemplary siRNAs for selective knockdown of MET full length (targeting exon junction 26 and 28 (SEQ ID Nos. 63 and 64) and variant (targeting exon 27 (SEQ ID Nos. 68 and 69)) are listed in Table 29. 
     The inventor also discovered a novel splicing variant of NF1 (Neurofibromin 1), which lacks exon 8 (SEQ ID No. 76, Table 31). The nucleotide sequence of NF1 full length cDNA sequence (SEQ ID No. 70) is shown in Table 30. Exon 8 is marked with double underline. Exemplary primer across the junction of splicing variant (SEQ ID No. 75) that is useful for detecting the presence of this variant is shown in Table 32. Exemplary siRNAs for selective knockdown of NF1 full length (targeting exon 8, SEQ ID NOs. 73 and 74) and variant (targeting exon junction (SEQ ID Nos. 77 and 78)) are listed in Table 33. 
     The inventor also discovered a novel splicing variant of BAK1 (Bcl-2 homologous antagonist/killer), which lacks exon 2 (SEQ ID No. 85, Table 35). The nucleotide sequence of BAK1 full length cDNA sequence (SEQ ID No. 79) is shown in Table 34. Exon 2 is marked with double underline. Exemplary primer across the junction of splicing variant (SEQ ID No. 84) that is useful for detecting the presence of this variant is shown in Table 36. Exemplary siRNAs for selective knockdown of BAK1 full length (targeting exon 2, SEQ ID NOs. 82 and 83) and variant (targeting exon junction (SEQ ID Nos. 86 and 87) are listed in Table 37. 
     
       
         
               
             
               
             
           
               
                 TABLE 2  
               
               
                   
               
               
                 PIK3CD (Full length)Nucleotide Sequence (3135 nt, SEQ ID 
               
               
                 No. 1) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGCCCCCTGGGGTGGACTGCCCCATGGAATTCTGGACCAAGGAGGAGAATCAGAGCGTTGTGGTTGACT 
               
               
                   
               
               
                 TCCTGCTGCCCACAGGGGTCTACCTGAACTTCCCTGTGTCCCGCAATGCCAACCTCAGCACCATCAAGCA 
               
               
                   
               
               
                 GCTGCTGTGGCACCGCGCCCAGTATGAGCCGCTCTTCCACATGCTCAGTGGCCCCGAGGCCTATGTGTTC 
               
               
                   
               
               
                 ACCTGCATCAACCAGACAGCGGAGCAGCAAGAGCTGGAGGACGAGCAACGGCGTCTGTGTGACGTGCAGC 
               
               
                   
               
               
                 CCTTCCTGCCCGTCCTGCGCCTGGTGGCCCGTGAGGGCGACCGCGTGAAGAAGCTCATCAACTCACAGAT 
               
               
                   
               
               
                 CAGCCTCCTCATCGGCAAAGGCCTCCACGAGTTTGACTCCTTGTGCGACCCAGAAGTGAACGACTTTCGC 
               
               
                   
               
               
                 GCCAAGATGTGCCAATTCTGCGAGGAGGCGGCCGCCCGCCGGCAGCAGCTGGGCTGGGAGGCCTGGCTGC 
               
               
                   
               
               
                 AGTACAGTTTCCCCCTGCAGCTGGAGCCCTCGGCTCAAACCTGGGGGCCTGGTACCCTGCGGCTCCCGAA 
               
               
                   
               
               
                 CCGGGCCCTTCTGGTCAACGTTAAGTTTGAGGGCAGCGAGGAGAGCTTCACCTTCCAGGTGTCCACCAAG 
               
               
                   
               
               
                 GACGTGCCGCTGGCGCTGATGGCCTGTGCCCTGCGGAAGAAGGCCACAGTGTTCCGGCAGCCGCTGGTGG 
               
               
                   
               
               
                 AGCAGCCGGAAGACTACACGCTGCAGGTGAACGGCAGGCATGAGTACCTGTATGGCAGCTACCCGCTCTG 
               
               
                   
               
               
                 CCAGTTCCAGTACATCTGCAGCTGCCTGCACAGTGGGTTGACCCCTCACCTGACCATGGTCCATTCCTCC 
               
               
                   
               
               
                 TCCATCCTCGCCATGCGGGATGAGCAGAGCAACCCTGCCCCCCAGGTCCAGAAACCGCGTGCCAAACCAC 
               
               
                   
               
               
                 CTCCCATTCCTGCGAAGAAG CCTTCCTCTGTGTCCCTGTGGTCCCTGGAGCAGCCGTTCCGCATCGAGCT   
               
               
                   
               
               
                   CATCCAGGGCAGCAAAGTGAACGCCGACGAGCGGATGAAG CTGGTGGTGCAGGCCGGGCTTTTCCACGGC 
               
               
                   
               
               
                 AACGAGATGCTGTGCAAGACGGTGTCCAGCTCGGAGGTGAGCGTGTGCTCGGAGCCCGTGTGGAAGCAGC 
               
               
                   
               
               
                 GGCTGGAGTTCGACATCAACATCTGCGACCTGCCCCGCATGGCCCGTCTCTGCTTTGCGCTGTACGCCGT 
               
               
                   
               
               
                 GATCGAGAAAGCCAAGAAGGCTCGCTCCACCAAGAAGAAGTCCAAGAAGGCGGACTGCCCCATTGCCTGG 
               
               
                   
               
               
                 GCCAACCTCATGCTGTTTGACTACAAGGACCAGCTTAAGACCGGGGAACGCTGCCTCTACATGTGGCCCT 
               
               
                   
               
               
                 CCGTCCCAGATGAGAAGGGCGAGCTGCTGAACCCCACGGGCACTGTGCGCAGTAACCCCAACACGGATAG 
               
               
                   
               
               
                 CGCCGCTGCCCTGCTCATCTGCCTGCCCGAGGTGGCCCCGCACCCCGTGTACTACCCCGCCCTGGAGAAG 
               
               
                   
               
               
                 ATCTTGGAGCTGGGGCGACACAGCGAGTGTGTGCATGTCACCGAGGAGGAGCAGCTGCAGCTGCGGGAAA 
               
               
                   
               
               
                 TCCTGGAGCGGCGGGGGTCTGGGGAGCTGTATGAGCACGAGAAGGACCTGGTGTGGAAGCTGCGGCATGA 
               
               
                   
               
               
                 AGTCCAGGAGCACTTCCCGGAGGCGCTAGCCCGGCTGCTGCTGGTCACCAAGTGGAACAAGCATGAGGAT 
               
               
                   
               
               
                 GTGGCCCAGATGCTCTACCTGCTGTGCTCCTGGCCGGAGCTGCCCGTCCTGAGCGCCCTGGAGCTGCTAG 
               
               
                   
               
               
                 ACTTCAGCTTCCCCGATTGCCACGTAGGCTCCTTCGCCATCAAGTCGCTGCGGAAACTGACGGACGATGA 
               
               
                   
               
               
                 GCTGTTCCAGTACCTGCTGCAGCTGGTGCAGGTGCTCAAGTACGAGTCCTACCTGGACTGCGAGCTGACC 
               
               
                   
               
               
                 AAATTCCTGCTGGACCGGGCCCTGGCCAACCGCAAGATCGGCCACTTCCTTTTCTGGCACCTCCGCTCCG 
               
               
                   
               
               
                 AGATGCACGTGCCGTCGGTGGCCCTGCGCTTCGGCCTCATCCTGGAGGCCTACTGCAGGGGCAGCACCCA 
               
               
                   
               
               
                 CCACATGAAGGTGCTGATGAAGCAGGGGGAAGCACTGAG   CAAACTGAAGGCCCTGAATGA   CTTCGTCAAG 
               
               
                   
               
               
                 CTGAGCTCTCAGAAGACCCCCAAGCCCCAGACCAAGGAGCTGATGCACTTGTGCATGCGGCAGGAGGCCT 
               
               
                   
               
               
                 ACCTAGAGGCCCTCTCCCACCTGCAGTCCCCACTCGACCCCAGCACCCTGCTGGCTGAAGTCTGCGTGGA 
               
               
                   
               
               
                 GCAGTGCACCTTCATGGACTCCAAGATGAAGCCCCTGTGGATCATGTACAGCAACGAGGAGGCAGGCAGC 
               
               
                   
               
               
                 GGCGGCAGCGTGGGCATCATCTTTAAGAACGGGGATGACCTCCGGCAGGACATGCTGACCCTGCAGATGA 
               
               
                   
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                 TGTGCTGGGCATTGGCGATCGGCACAG   CGACAACATCATGATCCGAGA   GAGTGGGCAGCTGTTCCACATT 
               
               
                   
               
               
                 GATTTTGGCCACTTTCTGGGGAATTTCAAGACCAAGTTTGGAATCAACCGCGAGCGTGTCCCATTCATCC 
               
               
                   
               
               
                 TCACCTACGACTTTGTCCATGTGATTCAGCGGGGAAGACTAATAATAGTTGAGAAATTTGAACGGTTCCG 
               
               
                   
               
               
                 GGGCTACTGTGAAAGGGCCTACACCATCCTGCGGCGCCACGGGCTTCTCTTCCTCCACCTCTTTGCCCTG 
               
               
                   
               
               
                 ATGCGGGCGGCAGGCCTGCCTGAGCTCAGCTGCTCCAAAGACATCCAGTATCTCAAGGACTCCCTGGCAC 
               
               
                   
               
               
                 TGGGGAAAACAGAGGAGGAGGCACTGAAGCACTTCCGAGTGAAGTTTAACGAAGCCCTCCGTGAGAGCTG 
               
               
                   
               
               
                 GAAAACCAAAGTGAACTGGCTGGCCCACAACGTGTCCAAAGACAACAGGCAGTAG 
               
               
                   
               
               
                 (Exon 10 is indicated by double underline, Exon 23 is indicated by wave underline. Primers for qRT-PCR validations of PIK3CD splice variants (-L and -S forms) are underlined) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 3 
               
               
                   
               
               
                 PIK3CD variant 1 (lacking exon 10) Nucleotide Sequence 
               
               
                 (3045 nt, SEQ ID No. 7) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGCCCCCTGGGGTGGACTGCCCCATGGAATTCTGGACCAAGGAGGAGAATCAGAGCGTTGTGGTTGACT 
               
               
                   
               
               
                 TCCTGCTGCCCACAGGGGTCTACCTGAACTTCCCTGTGTCCCGCAATGCCAACCTCAGCACCATCAAGCA 
               
               
                   
               
               
                 GCTGCTGTGGCACCGCGCCCAGTATGAGCCGCTCTTCCACATGCTCAGTGGCCCCGAGGCCTATGTGTTC 
               
               
                   
               
               
                 ACCTGCATCAACCAGACAGCGGAGCAGCAAGAGCTGGAGGACGAGCAACGGCGTCTGTGTGACGTGCAGC 
               
               
                   
               
               
                 CCTTCCTGCCCGTCCTGCGCCTGGTGGCCCGTGAGGGCGACCGCGTGAAGAAGCTCATCAACTCACAGAT 
               
               
                   
               
               
                 CAGCCTCCTCATCGGCAAAGGCCTCCACGAGTTTGACTCCTTGTGCGACCCAGAAGTGAACGACTTTCGC 
               
               
                   
               
               
                 GCCAAGATGTGCCAATTCTGCGAGGAGGCGGCCGCCCGCCGGCAGCAGCTGGGCTGGGAGGCCTGGCTGC 
               
               
                   
               
               
                 AGTACAGTTTCCCCCTGCAGCTGGAGCCCTCGGCTCAAACCTGGGGGCCTGGTACCCTGCGGCTCCCGAA 
               
               
                   
               
               
                 CCGGGCCCTTCTGGTCAACGTTAAGTTTGAGGGCAGCGAGGAGAGCTTCACCTTCCAGGTGTCCACCAAG 
               
               
                   
               
               
                 GACGTGCCGCTGGCGCTGATGGCCTGTGCCCTGCGGAAGAAGGCCACAGTGTTCCGGCAGCCGCTGGTGG 
               
               
                   
               
               
                 AGCAGCCGGAAGACTACACGCTGCAGGTGAACGGCAGGCATGAGTACCTGTATGGCAGCTACCCGCTCTG 
               
               
                   
               
               
                 CCAGTTCCAGTACATCTGCAGCTGCCTGCACAGTGGGTTGACCCCTCACCTGACCATGGTCCATTCCTCC 
               
               
                   
               
               
                 TCCATCCTCGCCATGCGGGATGAGCAGAGCAACCCTGCCCCCCAGGTCCAGAAACCGCGTGCCAAACCAC 
               
               
                   
               
               
                 CTCCCATTCC   TGCGAAGAA     GC     TGGTGGTGC   AGGCCGGGCTTTTCCACGGCAACGAGATGCTGTGCAAGACGG 
               
               
                   
               
               
                 TGTCCAGCTCGGAGGTGAGCGTGTGCTCGGAGCCCGTGTGGAAGCAGCGGCTGGAGTTCGACATCAACATCT 
               
               
                   
               
               
                 GCGACCTGCCCCGCATGGCCCGTCTCTGCTTTGCGCTGTACGCCGTGATCGAGAAAGCCAAGAAGGCTCGCT 
               
               
                   
               
               
                 CCACCAAGAAGAAGTCCAAGAAGGCGGACTGCCCCATTGCCTGGGCCAACCTCATGCTGTTTGACTACAAGG 
               
               
                   
               
               
                 ACCAGCTTAAGACCGGGGAACGCTGCCTCTACATGTGGCCCTCCGTCCCAGATGAGAAGGGCGAGCTGCTGA 
               
               
                   
               
               
                 ACCCCACGGGCACTGTGCGCAGTAACCCCAACACGGATAGCGCCGCTGCCCTGCTCATCTGCCTGCCCGAGG 
               
               
                   
               
               
                 TGGCCCCGCACCCCGTGTACTACCCCGCCCTGGAGAAGATCTTGGAGCTGGGGCGACACAGCGAGTGTGTGC 
               
               
                   
               
               
                 ATGTCACCGAGGAGGAGCAGCTGCAGCTGCGGGAAATCCTGGAGCGGCGGGGGTCTGGGGAGCTGTATGAGC 
               
               
                   
               
               
                 ACGAGAAGGACCTGGTGTGGAAGCTGCGGCATGAAGTCCAGGAGCACTTCCCGGAGGCGCTAGCCCGGCTGC 
               
               
                   
               
               
                 TGCTGGTCACCAAGTGGAACAAGCATGAGGATGTGGCCCAGATGCTCTACCTGCTGTGCTCCTGGCCGGAGC 
               
               
                   
               
               
                 TGCCCGTCCTGAGCGCCCTGGAGCTGCTAGACTTCAGCTTCCCCGATTGCCACGTAGGCTCCTTCGCCATCA 
               
               
                   
               
               
                 AGTCGCTGCGGAAACTGACGGACGATGAGCTGTTCCAGTACCTGCTGCAGCTGGTGCAGGTGCTCAAGTACG 
               
               
                   
               
               
                 AGTCCTACCTGGACTGCGAGCTGACCAAATTCCTGCTGGACCGGGCCCTGGCCAACCGCAAGATCGGCCACT 
               
               
                   
               
               
                 TCCTTTTCTGGCACCTCCGCTCCGAGATGCACGTGCCGTCGGTGGCCCTGCGCTTCGGCCTCATCCTGGAGG 
               
               
                   
               
               
                 CCTACTGCAGGGGCAGCACCCACCACATGAAGGTGCTGATGAAGCAGGGGGAAGCACTGAGCAAACTGAAGG 
               
               
                   
               
               
                 CCCTGAATGACTTCGTCAAGCTGAGCTCTCAGAAGACCCCCAAGCCCCAGACCAAGGAGCTGATGCACTTGT 
               
               
                   
               
               
                 GCATGCGGCAGGAGGCCTACCTAGAGGCCCTCTCCCACCTGCAGTCCCCACTCGACCCCAGCACCCTGCTGG 
               
               
                   
               
               
                 CTGAAGTCTGCGTGGAGCAGTGCACCTTCATGGACTCCAAGATGAAGCCCCTGTGGATCATGTACAGCAACG 
               
               
                   
               
               
                 AGGAGGCAGGCAGCGGCGGCAGCGTGGGCATCATCTTTAAGAACGGGGATGACCTCCGGCAGGACATGCTGA 
               
               
                   
               
               
                 CCCTGCAGATGATCCAGCTCATGGACGTCCTGTGGAAGCAGGAGGGGCTGGACCTGAGGATGACCCCCTATG 
               
               
                   
               
               
                 GCTGCCTCCCCACCGGGGACCGCACAGGCCTCATTGAGGTGGTACTCCGTTCAGACACCATCGCCAACATCC 
               
               
                   
               
               
                 AACTCAACAAGAGCAACATGGCAGCCACAGCCGCCTTCAACAAGGATGCCCTGCTCAACTGGCTGAAGTCCA 
               
               
                   
               
               
                 AGAACCCGGGGGAGGCCCTGGATCGAGCCATTGAGGAGTTCACCCTCTCCTGTGCTGGCTATTGTGTGGCCA 
               
               
                   
               
               
                 CATATGTGCTGGGCATTGGCGATCGGCACAGCGACAACATCATGATCCGAGAGAGTGGGCAGCTGTTCCACA 
               
               
                   
               
               
                 TTGATTTTGGCCACTTTCTGGGGAATTTCAAGACCAAGTTTGGAATCAACCGCGAGCGTGTCCCATTCATCC 
               
               
                   
               
               
                 TCACCTACGACTTTGTCCATGTGATTCAGCAGGGGAAGACTAATAATAGTGAGAAATTTGAACGGTTCCGGG 
               
               
                   
               
               
                 GCTACTGTGAAAGGGCCTACACCATCCTGCGGCGCCACGGGCTTCTCTTCCTCCACCTCTTTGCCCTGATGC 
               
               
                   
               
               
                 GGGCGGCAGGCCTGCCTGAGCTCAGCTGCTCCAAAGACATCCAGTATCTCAAGGACTCCCTGGCACTGGGGA 
               
               
                   
               
               
                 AAACAGAGGAGGAGGCACTGAAGCACTTCCGAGTGAAGTTTAACGAAGCCCTCCGTGAGAGCTGGAAAACCA 
               
               
                   
               
               
                 AAGTGAACTGGCTGGCCCACAACGTGTCCAAAGACAACAGGCAGTAG 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 4 
               
               
                   
               
               
                 PIK3CD variant 2 (lacking exon 23)Nucleotide Sequence 
               
               
                 (2967 nt, SEQ ID No. 11) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGCCCCCTGGGGTGGACTGCCCCATGGAATTCTGGACCAAGGAGGAGAATCAGAGCGTTGTGGTTGACT 
               
               
                   
               
               
                 TCCTGCTGCCCACAGGGGTCTACCTGAACTTCCCTGTGTCCCGCAATGCCAACCTCAGCACCATCAAGCA 
               
               
                   
               
               
                 GCTGCTGTGGCACCGCGCCCAGTATGAGCCGCTCTTCCACATGCTCAGTGGCCCCGAGGCCTATGTGTTC 
               
               
                   
               
               
                 ACCTGCATCAACCAGACAGCGGAGCAGCAAGAGCTGGAGGACGAGCAACGGCGTCTGTGTGACGTGCAGC 
               
               
                   
               
               
                 CCTTCCTGCCCGTCCTGCGCCTGGTGGCCCGTGAGGGCGACCGCGTGAAGAAGCTCATCAACTCACAGAT 
               
               
                   
               
               
                 CAGCCTCCTCATCGGCAAAGGCCTCCACGAGTTTGACTCCTTGTGCGACCCAGAAGTGAACGACTTTCGC 
               
               
                   
               
               
                 GCCAAGATGTGCCAATTCTGCGAGGAGGCGGCCGCCCGCCGGCAGCAGCTGGGCTGGGAGGCCTGGCTGC 
               
               
                   
               
               
                 AGTACAGTTTCCCCCTGCAGCTGGAGCCCTCGGCTCAAACCTGGGGGCCTGGTACCCTGCGGCTCCCGAA 
               
               
                   
               
               
                 CCGGGCCCTTCTGGTCAACGTTAAGTTTGAGGGCAGCGAGGAGAGCTTCACCTTCCAGGTGTCCACCAAG 
               
               
                   
               
               
                 GACGTGCCGCTGGCGCTGATGGCCTGTGCCCTGCGGAAGAAGGCCACAGTGTTCCGGCAGCCGCTGGTGG 
               
               
                   
               
               
                 AGCAGCCGGAAGACTACACGCTGCAGGTGAACGGCAGGCATGAGTACCTGTATGGCAGCTACCCGCTCTG 
               
               
                   
               
               
                 CCAGTTCCAGTACATCTGCAGCTGCCTGCACAGTGGGTTGACCCCTCACCTGACCATGGTCCATTCCTCC 
               
               
                   
               
               
                 TCCATCCTCGCCATGCGGGATGAGCAGAGCAACCCTGCCCCCCAGGTCCAGAAACCGCGTGCCAAACCAC 
               
               
                   
               
               
                 CTCCCATTCCTGCGAAGAAGCCTTCCTCTGTGTCCCTGTGGTCCCTGGAGCAGCCGTTCCGCATCGAGCT 
               
               
                   
               
               
                 CATCCAGGGCAGCAAAGTGAACGCCGACGAGCGGATGAAGCTGGTGGTGCAGGCCGGGCTTTTCCACGGC 
               
               
                   
               
               
                 AACGAGATGCTGTGCAAGACGGTGTCCAGCTCGGAGGTGAGCGTGTGCTCGGAGCCCGTGTGGAAGCAGC 
               
               
                   
               
               
                 GGCTGGAGTTCGACATCAACATCTGCGACCTGCCCCGCATGGCCCGTCTCTGCTTTGCGCTGTACGCCGT 
               
               
                   
               
               
                 GATCGAGAAAGCCAAGAAGGCTCGCTCCACCAAGAAGAAGTCCAAGAAGGCGGACTGCCCCATTGCCTGG 
               
               
                   
               
               
                 GCCAACCTCATGCTGTTTGACTACAAGGACCAGCTTAAGACCGGGGAACGCTGCCTCTACATGTGGCCCT 
               
               
                   
               
               
                 CCGTCCCAGATGAGAAGGGCGAGCTGCTGAACCCCACGGGCACTGTGCGCAGTAACCCCAACACGGATAG 
               
               
                   
               
               
                 CGCCGCTGCCCTGCTCATCTGCCTGCCCGAGGTGGCCCCGCACCCCGTGTACTACCCCGCCCTGGAGAAG 
               
               
                   
               
               
                 ATCTTGGAGCTGGGGCGACACAGCGAGTGTGTGCATGTCACCGAGGAGGAGCAGCTGCAGCTGCGGGAAA 
               
               
                   
               
               
                 TCCTGGAGCGGCGGGGGTCTGGGGAGCTGTATGAGCACGAGAAGGACCTGGTGTGGAAGCTGCGGCATGA 
               
               
                   
               
               
                 AGTCCAGGAGCACTTCCCGGAGGCGCTAGCCCGGCTGCTGCTGGTCACCAAGTGGAACAAGCATGAGGAT 
               
               
                   
               
               
                 GTGGCCCAGATGCTCTACCTGCTGTGCTCCTGGCCGGAGCTGCCCGTCCTGAGCGCCCTGGAGCTGCTAG 
               
               
                   
               
               
                 ACTTCAGCTTCCCCGATTGCCACGTAGGCTCCTTCGCCATCAAGTCGCTGCGGAAACTGACGGACGATGA 
               
               
                   
               
               
                 GCTGTTCCAGTACCTGCTGCAGCTGGTGCAGGTGCTCAAGTACGAGTCCTACCTGGACTGCGAGCTGACC 
               
               
                   
               
               
                 AAATTCCTGCTGGACCGGGCCCTGGCCAACCGCAAGATCGGCCACTTCCTTTTCTGGCACCTCCGCTCCG 
               
               
                   
               
               
                 AGATGCACGTGCCGTCGGTGGCCCTGCGCTTCGGCCTCATCCTGGAGGCCTACTGCAGGGGCAGCACCCA 
               
               
                   
               
               
                 CCACATGAAGGTGCTGATGAAGCAGGGGGAAGCACTGAGCAAACTGAAGGCCCTGAATGACTTCGTCAAG 
               
               
                   
               
               
                 CTGAGCTCTCAGAAGACCCCCAAGCCCCAGACCAAGGAGCTGATGCACTTGTGCATGCGGCAGGAGGCCT 
               
               
                   
               
               
                 ACCTAGAGGCCCTCTCCCACCTGCAGTCCCCACTCGACCCCAGCACCCTGCTGGCTGAAGTCTGCGTGGA 
               
               
                   
               
               
                 GCAGTGCACCTTCATGGACTCCAAGATGAAGCCCCTGTGGATCATGTACAGCAACGAGGAGGCAGGCAGC 
               
               
                   
               
               
                 GGCGGCAGCGTGGGCATCATCTTTAAGAACGGGGATGACCTCCGGCAGGACATGCTGACCCTGCAGATGA 
               
               
                   
               
               
                 TCCAGCTCATGGACGTCCTGTGGAAGCAGGAGGGGC   TGGACCTGA       GG       GAGGCCCT   GGATCGAGCCATTGAGG 
               
               
                   
               
               
                 AGTTCACCCTCTCCTGTGCTGGCTATTGTGTGGCCACATATGTGCTGGGCATTGGCGATCGGCACAGCGACA 
               
               
                   
               
               
                 ACATCATGATCCGAGAGAGTGGGCAGCTGTTCCACATTGATTTTGGCCACTTTCTGGGGAATTTCAAGACCA 
               
               
                   
               
               
                 AGTTTGGAATCAACCGCGAGCGTGTCCCATTCATCCTCACCTACGACTTTGTCCATGTGATTCAGCAGGGGA 
               
               
                   
               
               
                 AGACTAATAATAGTGAGAAATTTGAACGGTTCCGGGGCTACTGTGAAAGGGCCTACACCATCCTGCGGCGCC 
               
               
                   
               
               
                 ACGGGCTTCTCTTCCTCCACCTCTTTGCCCTGATGCGGGCGGCAGGCCTGCCTGAGCTCAGCTGCTCCAAAG 
               
               
                   
               
               
                 ACATCCAGTATCTCAAGGACTCCCTGGCACTGGGGAAAACAGAGGAGGAGGCACTGAAGCACTTCCGAGTGA 
               
               
                   
               
               
                 AGTTTAACGAAGCCCTCCGTGAGAGCTGGAAAACCAAAGTGAACTGGCTGGCCCACAACGTGTCCAAAGACA 
               
               
                   
               
               
                 ACAGGCAGTAG 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 5 
               
               
                   
               
               
                 PIK3CD variant 3 (lacking exon 10 and exon 23) 
               
               
                 Nucleotide Sequence (2877 nt, SEQ ID No. 14): 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGCCCCCTGGGGTGGACTGCCCCATGGAATTCTGGACCAAGGAGGAGAATCAGAGCGTTGTGGTTGACT 
               
               
                   
               
               
                 TCCTGCTGCCCACAGGGGTCTACCTGAACTTCCCTGTGTCCCGCAATGCCAACCTCAGCACCATCAAGCA 
               
               
                   
               
               
                 GCTGCTGTGGCACCGCGCCCAGTATGAGCCGCTCTTCCACATGCTCAGTGGCCCCGAGGCCTATGTGTTC 
               
               
                   
               
               
                 ACCTGCATCAACCAGACAGCGGAGCAGCAAGAGCTGGAGGACGAGCAACGGCGTCTGTGTGACGTGCAGC 
               
               
                   
               
               
                 CCTTCCTGCCCGTCCTGCGCCTGGTGGCCCGTGAGGGCGACCGCGTGAAGAAGCTCATCAACTCACAGAT 
               
               
                   
               
               
                 CAGCCTCCTCATCGGCAAAGGCCTCCACGAGTTTGACTCCTTGTGCGACCCAGAAGTGAACGACTTTCGC 
               
               
                   
               
               
                 GCCAAGATGTGCCAATTCTGCGAGGAGGCGGCCGCCCGCCGGCAGCAGCTGGGCTGGGAGGCCTGGCTGC 
               
               
                   
               
               
                 AGTACAGTTTCCCCCTGCAGCTGGAGCCCTCGGCTCAAACCTGGGGGCCTGGTACCCTGCGGCTCCCGAA 
               
               
                   
               
               
                 CCGGGCCCTTCTGGTCAACGTTAAGTTTGAGGGCAGCGAGGAGAGCTTCACCTTCCAGGTGTCCACCAAG 
               
               
                   
               
               
                 GACGTGCCGCTGGCGCTGATGGCCTGTGCCCTGCGGAAGAAGGCCACAGTGTTCCGGCAGCCGCTGGTGG 
               
               
                   
               
               
                 AGCAGCCGGAAGACTACACGCTGCAGGTGAACGGCAGGCATGAGTACCTGTATGGCAGCTACCCGCTCTG 
               
               
                   
               
               
                 CCAGTTCCAGTACATCTGCAGCTGCCTGCACAGTGGGTTGACCCCTCACCTGACCATGGTCCATTCCTCC 
               
               
                   
               
               
                 TCCATCCTCGCCATGCGGGATGAGCAGAGCAACCCTGCCCCCCAGGTCCAGAAACCGCGTGCCAAACCAC 
               
               
                   
               
               
                 CTCCCATTCC   TGCGAAGAA       GC       TGGTGGTG C AGGCCGGGCTTTTCCACGGCAACGAGATGCTGTGCAAGACGG 
               
               
                   
               
               
                 TGTCCAGCTCGGAGGTGAGCGTGTGCTCGGAGCCCGTGTGGAAGCAGCGGCTGGAGTTCGACATCAACATCT 
               
               
                   
               
               
                 GCGACCTGCCCCGCATGGCCCGTCTCTGCTTTGCGCTGTACGCCGTGATCGAGAAAGCCAAGAAGGCTCGCT 
               
               
                   
               
               
                 CCACCAAGAAGAAGTCCAAGAAGGCGGACTGCCCCATTGCCTGGGCCAACCTCATGCTGTTTGACTACAAGG 
               
               
                   
               
               
                 ACCAGCTTAAGACCGGGGAACGCTGCCTCTACATGTGGCCCTCCGTCCCAGATGAGAAGGGCGAGCTGCTGA 
               
               
                   
               
               
                 ACCCCACGGGCACTGTGCGCAGTAACCCCAACACGGATAGCGCCGCTGCCCTGCTCATCTGCCTGCCCGAGG 
               
               
                   
               
               
                 TGGCCCCGCACCCCGTGTACTACCCCGCCCTGGAGAAGATCTTGGAGCTGGGGCGACACAGCGAGTGTGTGC 
               
               
                   
               
               
                 ATGTCACCGAGGAGGAGCAGCTGCAGCTGCGGGAAATCCTGGAGCGGCGGGGGTCTGGGGAGCTGTATGAGC 
               
               
                   
               
               
                 ACGAGAAGGACCTGGTGTGGAAGCTGCGGCATGAAGTCCAGGAGCACTTCCCGGAGGCGCTAGCCCGGCTGC 
               
               
                   
               
               
                 TGCTGGTCACCAAGTGGAACAAGCATGAGGATGTGGCCCAGATGCTCTACCTGCTGTGCTCCTGGCCGGAGC 
               
               
                   
               
               
                 TGCCCGTCCTGAGCGCCCTGGAGCTGCTAGACTTCAGCTTCCCCGATTGCCACGTAGGCTCCTTCGCCATCA 
               
               
                   
               
               
                 AGTCGCTGCGGAAACTGACGGACGATGAGCTGTTCCAGTACCTGCTGCAGCTGGTGCAGGTGCTCAAGTACG 
               
               
                   
               
               
                 AGTCCTACCTGGACTGCGAGCTGACCAAATTCCTGCTGGACCGGGCCCTGGCCAACCGCAAGATCGGCCACT 
               
               
                   
               
               
                 TCCTTTTCTGGCACCTCCGCTCCGAGATGCACGTGCCGTCGGTGGCCCTGCGCTTCGGCCTCATCCTGGAGG 
               
               
                   
               
               
                 CCTACTGCAGGGGCAGCACCCACCACATGAAGGTGCTGATGAAGCAGGGGGAAGCACTGAGCAAACTGAAGG 
               
               
                   
               
               
                 CCCTGAATGACTTCGTCAAGCTGAGCTCTCAGAAGACCCCCAAGCCCCAGACCAAGGAGCTGATGCACTTGT 
               
               
                   
               
               
                 GCATGCGGCAGGAGGCCTACCTAGAGGCCCTCTCCCACCTGCAGTCCCCACTCGACCCCAGCACCCTGCTGG 
               
               
                   
               
               
                 CTGAAGTCTGCGTGGAGCAGTGCACCTTCATGGACTCCAAGATGAAGCCCCTGTGGATCATGTACAGCAACG 
               
               
                   
               
               
                 AGGAGGCAGGCAGCGGCGGCAGCGTGGGCATCATCTTTAAGAACGGGGATGACCTCCGGCAGGACATGCTGA 
               
               
                   
               
               
                 CCCTGCAGATGATCCAGCTCATGGACGTCCTGTGGAAGCAGGAGGGGC   TGGACCTGA       GG       GAGGCCC   TGGATC 
               
               
                   
               
               
                 GAGCCATTGAGGAGTTCACCCTCTCCTGTGCTGGCTATTGTGTGGCCACATATGTGCTGGGCATTGGCGATC 
               
               
                   
               
               
                 GGCACAGCGACAACATCATGATCCGAGAGAGTGGGCAGCTGTTCCACATTGATTTTGGCCACTTTCTGGGGA 
               
               
                   
               
               
                 ATTTCAAGACCAAGTTTGGAATCAACCGCGAGCGTGTCCCATTCATCCTCACCTACGACTTTGTCCATGTGA 
               
               
                   
               
               
                 TTCAGCAGGGGAAGACTAATAATAGTGAGAAATTTGAACGGTTCCGGGGCTACTGTGAAAGGGCCTACACCA 
               
               
                   
               
               
                 TCCTGCGGCGCCACGGGCTTCTCTTCCTCCACCTCTTTGCCCTGATGCGGGCGGCAGGCCTGCCTGAGCTCA 
               
               
                   
               
               
                 GCTGCTCCAAAGACATCCAGTATCTCAAGGACTCCCTGGCACTGGGGAAAACAGAGGAGGAGGCACTGAAGC 
               
               
                   
               
               
                 ACTTCCGAGTGAAGTTTAACGAAGCCCTCCGTGAGAGCTGGAAAACCAAAGTGAACTGGCTGGCCCACAACG 
               
               
                   
               
               
                 TGTCCAAAGACAACAGGCAGTAG 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 6 
               
               
                   
               
               
                 PIK3CD variant 4 (with large deletion) Nucleotide 
               
               
                 Sequence (1836 nt, SEQ ID No. 16): 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGCCCCCTGGGGTGGACTGCCCCATGGAATTCTGGACCAAGGAGGAGAATCAGAGCGTTGTGGTTGACT 
               
               
                   
               
               
                 TCCTGCTGCCCACAGGGGTCTACCTGAACTTCCCTGTGTCCCGCAATGCCAACCTCAGCACCATCAAGCA 
               
               
                   
               
               
                 GCTGCTGTGGCACCGCGCCCAGTATGAGCCGCTCTTCCACATGCTCAGTGGCCCCGAGGCCTATGTGTTC 
               
               
                   
               
               
                 ACCTGCATCAACCAGACAGCGGAGCAGCAAGAGCTGGAGGACGAGCAACGGCGTCTGTGTGACGTGCAGC 
               
               
                   
               
               
                 CCTTCCTGCCCGTCCTGCGCCTGGTGGCCCGTGAGGGCGACCGCGTGAAGAAGCTCATCAACTCACAGAT 
               
               
                   
               
               
                 CAGCCTCCTCATCGGCAAAGGCCTCCACGAGTTTGACTCCTTGTGCGACCCAGAAGTGAACGACTTTCGC 
               
               
                   
               
               
                 GCCAAGATGTGCCAATTCTGCGAGGAGGCGGCCGCCCGCCGGCAGCAGCTGGGCTGGGAGGCCTGGCTGC 
               
               
                   
               
               
                 AGTACAGTTTCCCCCTGCAGCTGGAGCCCTCGGCTCAAACCTGGGGGCCTGGTACCCTGCGGCTCCCGAA 
               
               
                   
               
               
                 CCGGGCCCTTCTGGTCAACGTTAAGTTTGAGGGCAGCGAGGAGAGCTTCACCTTCCAGGTGTCCACCAAG 
               
               
                   
               
               
                 GACGTGCCGCTGGCGCTGATGGCCTGTGCCCTGCGGAAGAAGGCCACAGTGTTCCGGCAGCCGCTGGTGG 
               
               
                   
               
               
                 AGCAGCCGGAAGACTACACGCTGCAGGTGAACGGCAGGCATGAGTACCTGTATGGCAGCTACCCGCTCTG 
               
               
                   
               
               
                 CCAGTTCCAGTACATCTGCAGCTGCCTGCACAGTGGGTTGACCCCTCACCTGACCATGGTCCATTCCTCC 
               
               
                   
               
               
                 TCCATCCTCGCCATGCGGGATGAGCAGAGCAACCCTGCCCCCCAGGTCCAGAAACCGCGTGCCAAACCAC 
               
               
                   
               
               
                 CTCCCATTCCTGCGAAGAAGCCTTCCTCTGTGTCCCTGTGGTCCCTGGAGCAGCCGTTCCGCATCGAGCT 
               
               
                   
               
               
                 CATCCAGGGCAGCAAAGTGAACGCCGACGAGCGGATGAAGCTGGTGGTGCAGGCCGGGCTTTTCCACGGC 
               
               
                   
               
               
                 AACGAGATGCTGTGCAAGACGGTGTCCAGCTCGGAGGTGAGCGTGTGCTCGGAGCCCGTGTGGAAGCAGC 
               
               
                   
               
               
                 GGCTGGAGTTCGACATCAACATCTGCGACCTGCCCCGCATGGCCCGTCTCTGCTTTGCGCTGTACGCCGT 
               
               
                   
               
               
                 GATCGAGAAAGCCAAGAAGGCTCGCTCCACCAAGAAGAAGTCCAAGAAGGCGGACTGCCCCATTGCCTGG 
               
               
                   
               
               
                 GCCAACCTCATGCTGTTTGACTACAAGGACCAGCTTAAGACCGGGGAACGCTGCCTCT   ACATGTGGCC       CC       TC     
               
               
                   
               
               
                     TCCTGT   GCTGGCTATTGTGTGGCCACATATGTGCTGGGCATTGGCGATCGGCACAGCGACAACATCATGATC 
               
               
                   
               
               
                 CGAGAGAGTGGGCAGCTGTTCCACATTGATTTTGGCCACTTTCTGGGGAATTTCAAGACCAAGTTTGGAATC 
               
               
                   
               
               
                 AACCGCGAGCGTGTCCCATTCATCCTCACCTACGACTTTGTCCATGTGATTCAGCAGGGGAAGACTAATAAT 
               
               
                   
               
               
                 AGTGAGAAATTTGAACGGTTCCGGGGCTACTGTGAAAGGGCCTACACCATCCTGCGGCGCCACGGGCTTCTC 
               
               
                   
               
               
                 TTCCTCCACCTCTTTGCCCTGATGCGGGCGGCAGGCCTGCCTGAGCTCAGCTGCTCCAAAGACATCCAGTAT 
               
               
                   
               
               
                 CTCAAGGACTCCCTGGCACTGGGGAAAACAGAGGAGGAGGCACTGAAGCACTTCCGAGTGAAGTTTAACGAA 
               
               
                   
               
               
                 GCCCTCCGTGAGAGCTGGAAAACCAAAGTGAACTGGCTGGCCCACAACGTGTCCAAAGACAACAGGCAGTAG 
               
               
                   
               
               
                 (Double underline indicates bases bordering the deletion junction) 
               
             
          
         
       
     
     
       
         
               
             
               
               
             
           
               
                 TABLE 7 
               
               
                   
               
               
                 Primers for detecting PIK3CD variants 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Primer across the junction 
                 
                   TGCGAAGAA 
                   GC 
                   TGGTGGTGC 
                 
               
               
                 between PIK3CD exon 9 and 11 
                   
               
               
                 (SEQ ID No. 6) 
                   
               
               
                   
               
               
                 Primer sequences across the 
                 
                   
                     TGGACCTGA 
                   
                   
                     GG 
                   
                   
                     GAGGCCCT 
                   
                 
               
               
                 junct. between PIK3CD exon 
                   
               
               
                 22 and 24 (SEQ ID No. 10) 
                   
               
               
                   
               
               
                 Primer sequences across the 
                 
                   
                     ACATGTGGCC 
                   
                   
                     CC 
                   
                   
                     TCTCCTG 
                   
                 
               
               
                 deleted region (nt1329-2627) 
                   
               
               
                 of PIK3CD 
                   
               
               
                 (SEQ ID No. 15) : 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
           
               
                 TABLE 8 
               
               
                   
               
               
                 siRNA for selectively knockdown PIK3CD full 
               
               
                 length and variants expression 
               
               
                   
               
             
             
               
                 siRNA targeting PIK3CD exon 23 (siPIK3CD-ex23) 
               
               
                 Sense (SEQ ID No. 4): 5′ CCAACAUCCAACUCAACAAdTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 5): 3′ dTdTGGUUGUAGGUUGAGUU- 
               
               
                 GUU (5′-P)5′ 
               
               
                   
               
               
                 siRNA targeting junction spanning between 
               
               
                 exon 9 and exon 11 
               
               
                 Sense (SEQ ID No. 8) 5′ CUGCGAAGAA GC UGGUGGUdTdT 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 9) 3′ dTdTGACGCUUCUU CG ACCAC- 
               
               
                 CA (5′-P)5′ 
               
               
                   
               
               
                 siRNA targeting junction spanning between 
               
               
                 PIK3CD exon22 and exon 24 (siPIK3CD-S) 
               
               
                 Sense (SEQ ID No. 12) 5′ UGA GG GAGGCCCUGGAUCGAdTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 13) 3′ dTdTACU CC CUCCGGGACCU- 
               
               
                 AGCU (5′-P)5′ 
               
               
                   
               
               
                 siRNA targeting junction spanning the 
               
               
                 deleted sequences of PIK3CD variant 4 
               
               
                 Sense (SEQ ID No. 17) 5′  CC UCUCCUGUGCUGGCUAUdTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 18) 3′ dTdT GG AGAGGACACGACCG- 
               
               
                 AUA (5′-P)5′ 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 9 
               
               
                   
               
               
                 FGFR3 (Full length) Nucleotide Sequence 
               
               
                 (2421 nt, SEQ ID No. 19) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGGGCGCCCCTGCCTGCGCCCTCGCGCTCTGCGTGGCCGTGGCCATCGT 
               
               
                   
               
               
                 GGCCGGCGCCTCCTCGGAGTCCTTGGGGACGGAGCAGCGCGTCGTGGGGC 
               
               
                   
               
               
                 GAGCGGCAGAAGTCCCGGGCCCAGAGCCCGGCCAGCAGGAGCAGTTGGTC 
               
               
                   
               
               
                 TTCGGCAGCGGGGATGCTGTGGAGCTGAGCTGTCCCCCGCCCGGGGGTGG 
               
               
                   
               
               
                 TCCCATGGGGCCCACTGTCTGGGTCAAGGATGGCACAGGGCTGGTGCCCT 
               
               
                   
               
               
                 CGGAGCGTGTCCTGGTGGGGCCCCAGCGGCTGCAGGTGCTGAATGCCTCC 
               
               
                   
               
               
                 CACGAGGACTCCGGGGCCTACAGCTGCCGGCAGCGGCTCACGCAGCGCGT 
               
               
                   
               
               
                 ACTGTGCCACTTCAGTGTGCGGGTGACAGACGCTCCATCCTCGGGAGATG 
               
               
                   
               
               
                 ACGAAGACGGGGAGGACGAGGCTGAGGACACAGGTGTGGACACAGGGGCC 
               
               
                   
               
               
                 CCTTACTGGACACGGCCCGAGCGGATGGACAAGAAGCTGCTGGCCGTGCC 
               
               
                   
               
               
                 GGCCGCCAACACCGTCCGCTTCCGCTGCCCAGCCGCTGGCAACCCCACTC 
               
               
                   
               
               
                 CCTCCATCTCCTGGCTGAAGAACGGCAGGGAGTTCCGCGGCGAGCACCGC 
               
               
                   
               
               
                 ATTGGAGGCATCAAGCTGCGGCATCAGCAGTGGAGCCTGGTCATGGAAAG 
               
               
                   
               
               
                 CGTGGTGCCCTCGGACCGCGGCAACTACACCTGCGTCGTGGAGAACAAGT 
               
               
                   
               
               
                 TTGGCAGCATCCGGCAGACGTACACGCTGGACGTGCTGGAGCGCTCCCCG 
               
               
                   
               
               
                 CACCGGCCCATCCTGCAGGCGGGGCTGCCGGCCAACCAGACGGCGGTGCT 
               
               
                   
               
               
                 GGGCAGCGACGTGGAGTTCCACTGCAAGGTGTACAGTGACGCACAGCCCC 
               
               
                   
               
               
                 ACATCCAGTGGCTCAAGCACGTGGAGGTGAATGGCAGCAAGGTGGGCCCG 
               
               
                   
               
               
                 GACGGCACACCCTACGTTACCGTGCTCAAGACGGCGGGCGCTAACACCAC 
               
               
                   
               
               
                 CGACAAGGAGCTAGAGGTTCTCTCCTTGCACAACGTCACCTTTGAGGACG 
               
               
                   
               
               
                 CCGGGGAGTACACCTGCCTGGCGGGCAATTCTATTGGGTTTTCTCATCAC 
               
               
                   
               
               
                 TCTGCGTGGCTGGTGGTGCTGCCAGCCGAGGAGGAGCTGGTGGAGGCTGA 
               
               
                   
               
               
                 CGAGGCGGGCAGTGTGTATGCAGGCATCCTCAGCTACGGGGTGGGCTTCT 
               
               
                   
               
               
                 TCCTGTTCATCCTGGTGGTGGCGGCTGTGACGCTCTGCCGCCTGCGCAGC 
               
               
                   
               
               
                 CCCCCCAAGAAAGGCCTGGGCTCCCCCACCGTGCACAAGATCTCCCGCTT 
               
               
                   
               
               
                 CCCGCTCAAGCGACAGGTGTCCCTGGAGTCCAACGCGTCCATGAGCTCCA 
               
               
                   
               
               
                 ACACACCACTGGTGCGCATCGCAAGGCTGTCCTCAGGGGAGGGCCCCACG 
               
               
                   
               
               
                 CTGGCCAATGTCTCCGAGCTCGAGCTGCCTGCCGACCCCAAATGGGAGCT 
               
               
                   
               
               
                 GTCTCGGGCCCGGCTGACCCTGGGCAAGCCCCTTGGGGAGGGCTGCTTCG 
               
               
                   
               
               
                 GCCAGGTGGTCATGGCGGAGGCCATCGGCATTGACAAGGACCGGGCCGCC 
               
               
                   
               
               
                 AAGCCTGTCACCGTAGCCGTGAAGATGCTGAAAGACGATGCCACTGACAA 
               
               
                   
               
               
                 GGACCTGTCGGACCTGGTGTCTGAGATGGAGATGATGAAGATGATCGGGA 
               
               
                   
               
               
                 AACACAAAAACATCATCAACCTGCTGGGCGCCTGCACGCAGGGCGGGCCC 
               
               
                   
               
               
                 CTGTACGTGCTGGTGGAGTACGCGGCCAAGGGTAACCTGCGGGAGTTTCT 
               
               
                   
               
               
                 GCGGGCGCGGCGGCCCCCGGGCCTGGACTACTCCTTCGACACCTGCAAGC 
               
               
                   
               
               
                 CGCCCGAGGAGCAGCTCACCTTCAAGGACCTGGTGTCCTGTGCCTACCAG 
               
               
                   
               
               
                 GTGGCCCGGGGCATGGAGTACTTGGCCTCCCAGAAG TGCATCCACAGGGA   
               
               
                   
               
               
                 
                   CCTGGCTGCCCGCAATGTGCTGGTGACCGAGGACAACGTGATGAAGATCG 
                 
               
               
                   
               
               
                 
                   CAGACTTCGGGCTGGCCCGGGACGTGCACAACCTCGACTACTACAAGAAG 
                 
               
               
                   
               
               
                   ACGACCAAC GGCCGGCTGCCCGTGAAGTGGATGGCGCCTGAGGCCTTGTT 
               
               
                   
               
               
                 TGACCGAGTCTACACTCACCAGAGTGACGTCTGGTCCTTTGGGGTCCTGC 
               
               
                   
               
               
                 TCTGGGAGATCTTCACGCTGGGGGGCTCCCCGTACCCCGGCATCCCTGTG 
               
               
                   
               
               
                 GAGGAGCTCTTCAAGCTGCTGAAGGAGGGCCACCGCATGGACAAGCCCG   C     
               
               
                   
               
               
                     C   AACTGCACACACGACCT   GTACATGATCATGCGGGAGTGCTGGCATGCCG 
               
               
                   
               
               
                 CGCCCTCCCAGAGGCCCACCTTCAAGCAGCTGGTGGAGGACCTGGACCGT 
               
               
                   
               
               
                 GTCCTTACCGTGACGTCCACCGACGAGTACCTGGACCTGTCGGCGCCTTT 
               
               
                   
               
               
                 CGAGCAGTACTCCCCGGGTGGCCAGGACACCCCCAGCTCCAGCTCCTCAG 
               
               
                   
               
               
                 GGGACGACTCCGTGTTTGCCCACGACCTGCTGCCCCCGGCCCCACCCAGC 
               
               
                   
               
               
                 AGTGGGGGCTCGCGGACGTGA 
               
               
                   
               
               
                 (Exon 14 is indicated by double underline. Primers useful for detection of exon 14 splicing variants are underlined.) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 10 
               
               
                   
               
               
                 FGFR3 variant 1 (lacking exon 14) Nucleotide 
               
               
                 Sequence (2298 nt, SEQ ID No. 25): 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGGGCGCCCCTGCCTGCGCCCTCGCGCTCTGCGTGGCCGTGGCCATCGT 
               
               
                   
               
               
                 GGCCGGCGCCTCCTCGGAGTCCTTGGGGACGGAGCAGCGCGTCGTGGGGC 
               
               
                   
               
               
                 GAGCGGCAGAAGTCCCGGGCCCAGAGCCCGGCCAGCAGGAGCAGTTGGTC 
               
               
                   
               
               
                 TTCGGCAGCGGGGATGCTGTGGAGCTGAGCTGTCCCCCGCCCGGGGGTGG 
               
               
                   
               
               
                 TCCCATGGGGCCCACTGTCTGGGTCAAGGATGGCACAGGGCTGGTGCCCT 
               
               
                   
               
               
                 CGGAGCGTGTCCTGGTGGGGCCCCAGCGGCTGCAGGTGCTGAATGCCTCC 
               
               
                   
               
               
                 CACGAGGACTCCGGGGCCTACAGCTGCCGGCAGCGGCTCACGCAGCGCGT 
               
               
                   
               
               
                 ACTGTGCCACTTCAGTGTGCGGGTGACAGACGCTCCATCCTCGGGAGATG 
               
               
                   
               
               
                 ACGAAGACGGGGAGGACGAGGCTGAGGACACAGGTGTGGACACAGGGGCC 
               
               
                   
               
               
                 CCTTACTGGACACGGCCCGAGCGGATGGACAAGAAGCTGCTGGCCGTGCC 
               
               
                   
               
               
                 GGCCGCCAACACCGTCCGCTTCCGCTGCCCAGCCGCTGGCAACCCCACTC 
               
               
                   
               
               
                 CCTCCATCTCCTGGCTGAAGAACGGCAGGGAGTTCCGCGGCGAGCACCGC 
               
               
                   
               
               
                 ATTGGAGGCATCAAGCTGCGGCATCAGCAGTGGAGCCTGGTCATGGAAAG 
               
               
                   
               
               
                 CGTGGTGCCCTCGGACCGCGGCAACTACACCTGCGTCGTGGAGAACAAGT 
               
               
                   
               
               
                 TTGGCAGCATCCGGCAGACGTACACGCTGGACGTGCTGGAGCGCTCCCCG 
               
               
                   
               
               
                 CACCGGCCCATCCTGCAGGCGGGGCTGCCGGCCAACCAGACGGCGGTGCT 
               
               
                   
               
               
                 GGGCAGCGACGTGGAGTTCCACTGCAAGGTGTACAGTGACGCACAGCCCC 
               
               
                   
               
               
                 ACATCCAGTGGCTCAAGCACGTGGAGGTGAATGGCAGCAAGGTGGGCCCG 
               
               
                   
               
               
                 GACGGCACACCCTACGTTACCGTGCTCAAGACGGCGGGCGCTAACACCAC 
               
               
                   
               
               
                 CGACAAGGAGCTAGAGGTTCTCTCCTTGCACAACGTCACCTTTGAGGACG 
               
               
                   
               
               
                 CCGGGGAGTACACCTGCCTGGCGGGCAATTCTATTGGGTTTTCTCATCAC 
               
               
                   
               
               
                 TCTGCGTGGCTGGTGGTGCTGCCAGCCGAGGAGGAGCTGGTGGAGGCTGA 
               
               
                   
               
               
                 CGAGGCGGGCAGTGTGTATGCAGGCATCCTCAGCTACGGGGTGGGCTTCT 
               
               
                   
               
               
                 TCCTGTTCATCCTGGTGGTGGCGGCTGTGACGCTCTGCCGCCTGCGCAGC 
               
               
                   
               
               
                 CCCCCCAAGAAAGGCCTGGGCTCCCCCACCGTGCACAAGATCTCCCGCTT 
               
               
                   
               
               
                 CCCGCTCAAGCGACAGGTGTCCCTGGAGTCCAACGCGTCCATGAGCTCCA 
               
               
                   
               
               
                 ACACACCACTGGTGCGCATCGCAAGGCTGTCCTCAGGGGAGGGCCCCACG 
               
               
                   
               
               
                 CTGGCCAATGTCTCCGAGCTCGAGCTGCCTGCCGACCCCAAATGGGAGCT 
               
               
                   
               
               
                 GTCTCGGGCCCGGCTGACCCTGGGCAAGCCCCTTGGGGAGGGCTGCTTCG 
               
               
                   
               
               
                 GCCAGGTGGTCATGGCGGAGGCCATCGGCATTGACAAGGACCGGGCCGCC 
               
               
                   
               
               
                 AAGCCTGTCACCGTAGCCGTGAAGATGCTGAAAGACGATGCCACTGACAA 
               
               
                   
               
               
                 GGACCTGTCGGACCTGGTGTCTGAGATGGAGATGATGAAGATGATCGGGA 
               
               
                   
               
               
                 AACACAAAAACATCATCAACCTGCTGGGCGCCTGCACGCAGGGCGGGCCC 
               
               
                   
               
               
                 CTGTACGTGCTGGTGGAGTACGCGGCCAAGGGTAACCTGCGGGAGTTTCT 
               
               
                   
               
               
                 GCGGGCGCGGCGGCCCCCGGGCCTGGACTACTCCTTCGACACCTGCAAGC 
               
               
                   
               
               
                 CGCCCGAGGAGCAGCTCACCTTCAAGGACCTGGTGTCCTGTGCCTACCAG 
               
               
                   
               
               
                 GTGGCCCGGGGCATGGAGTAC TTGGCCTCCCAGAA   GG   GCCGGCT GCCCGT 
               
               
                   
               
               
                 GAAGTGGATGGCGCCTGAGGCCTTGTTTGACCGAGTCTACACTCACCAGA 
               
               
                   
               
               
                 GTGACGTCTGGTCCTTTGGGGTCCTGCTCTGGGAGATCTTCACGCTGGGG 
               
               
                   
               
               
                 GGCTCCCCGTACCCCGGCATCCCTGTGGAGGAGCTCTTCAAGCTGCTGAA 
               
               
                   
               
               
                 GGAGGGCCACCGCATGGACAAGCCCGCCAACTGCACACACGACCTGTACA 
               
               
                   
               
               
                 TGATCATGCGGGAGTGCTGGCATGCCGCGCCCTCCCAGAGGCCCACCTTC 
               
               
                   
               
               
                 AAGCAGCTGGTGGAGGACCTGGACCGTGTCCTTACCGTGACGTCCACCGA 
               
               
                   
               
               
                 CGAGTACCTGGACCTGTCGGCGCCTTTCGAGCAGTACTCCCCGGGTGGCC 
               
               
                   
               
               
                 AGGACACCCCCAGCTCCAGCTCCTCAGGGGACGACTCCGTGTTTGCCCAC 
               
               
                   
               
               
                 GACCTGCTGCCCCCGGCCCCACCCAGCAGTGGGGGCTCGCGGACGTGA 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
               
             
           
               
                 TABLE 11 
               
               
                   
               
               
                 Primer across the junction between 
               
               
                 FGFR3 exon 13 and 15 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Primer across the  
                 
                   TTGGCCTCCCAGAA 
                   GG 
                   GCCGGCT 
                 
               
               
                 junction between FGFR3  
                   
               
               
                 exon 13 and 15 
                   
               
               
                 (SEQ ID No. 24) 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
           
               
                 TABLE 12 
               
               
                   
               
               
                 siRNA for selectively knockdown FGFR3 
               
               
                 full length and variants expression 
               
               
                   
               
             
             
               
                 siRNA targeting FGFR3 exon 14: 
               
               
                 Sense (SEQ ID No. 22) 5′ CUCGACUACUACAAGAAGAdTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 23) 3′ dTdTGAGCUGAUGAUGUUCU- 
               
               
                 UCU (5′-P)5′ 
               
               
                   
               
               
                 siRNA targeting splice junction 
               
               
                 between FGFR3 exon 13 and 15 
               
               
                 Sense (SEQ ID No. 26) 5′  CCUCCCAGAA   GG   GCCGGCU  dTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 27) 3′ dTdTGGAGGGUCUU CC CGGC- 
               
               
                 CGA (5′-P)5′ 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 13 
               
               
                   
               
               
                 TSC2 (full length)Nucleotide Sequence (5424 nt, SEQ ID No. 28) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGGCCAAACCAACAAGCAAAGATTCAGGCTTGAAGGAGAAGTTTAAGATTCTGTTGGGACTGGGAACACCG 
               
               
                   
               
               
                 AGGCCAAATCCCAGGTCTGCAGAGGGTAAACAGACGGAGTTTATCATCACCGCGGAAATACTGAGAGAACTG 
               
               
                   
               
               
                 AGCATGGAATGTGGCCTCAACAATCGCATCCGGATGATAGGGCAGATTTGTGAAGTCGCAAAAACCAAGAAA 
               
               
                   
               
               
                 TTTGAAGAGCACGCAGTGGAAGCACTCTGGAAGGCGGTCGCGGATCTGTTGCAGCCGGAGCGGCCGCTGGAG 
               
               
                   
               
               
                 GCCCGGCACGCGGTGCTGGCTCTGCTGAAGGCCATCGTGCAGGGGCAGGGCGAGCGTTTGGGGGTCCTCAGA 
               
               
                   
               
               
                 GCCCTCTTCTTTAAGGTCATCAAGGATTACCCTTCCAACGAAGACCTTCACGAAAGGCTGGAGGTTTTCAAG 
               
               
                   
               
               
                 GCCCTCACAGACAATGGGAGACACATCACCTACTTGGAGGAAGAGCTGGCTGACTTTGTCCTGCAGTGGATG 
               
               
                   
               
               
                 GATGTTGGCTTGTCCTCGGAATTCCTTCTGGTGCTGGTGAACTTGGTCAAATTCAATAGCTGTTACCTCGAC 
               
               
                   
               
               
                 GAGTACATCGCAAGGATGGTTCAGATGATCTGTCTGCTGTGCGTCCGGACCGCGTCCTCTGTGGACATAGAG 
               
               
                   
               
               
                 GTCTCCCTGCAGGTGCTGGACGCCGTGGTCTGCTACAACTGCCTGCCGGCTGAGAGCCTCCCGCTGTTCATC 
               
               
                   
               
               
                 GTTACCCTCTGTCGCACCATCAACGTCAAGGAGCTCTGCGAGCCTTGCTGGAAGCTGATGCGGAACCTCCTT 
               
               
                   
               
               
                 GGCACCCACCTGGGCCACAGCGCCATCTACAACATGTGCCACCTCATGGAGGACAGAGCCTACATGGAGGAC 
               
               
                   
               
               
                 GCGCCCCTGCTGAGAGGAGCCGTGTTTTTTGTGGGCATGGCTCTCTGGGGAGCCCACCGGCTCTATTCTCTC 
               
               
                   
               
               
                 AGGAACTCGCCGACATCTGTGTTGCCATCATTTTACCAGGCCATGGCATGTCCGAACGAGGTGGTGTCCTAT 
               
               
                   
               
               
                 GAGATCGTCCTGTCCATCACCAGGCTCATCAAGAAGTATAGGAAGGAGCTCCAGGTGGTGGCGTGGGACATT 
               
               
                   
               
               
                 CTGCTGAACATCATCGAACGGCTCCTTCAGCAGCTCCAGACCTTGGACAGCCCGGAGCTCAGGACCATCGTC 
               
               
                   
               
               
                 CATGACCTGTTGACCACGGTGGAGGAGCTGTGTGACCAGAACGAGTTCCACGGGTCTCAGGAGAGATACTTT 
               
               
                   
               
               
                 GAACTGGTGGAGAGATGTGCGGACCAGAGGCCTGAGTCCTCCCTCCTGAACCTGATCTCCTATAGAGCGCAG 
               
               
                   
               
               
                 TCCATCCACCCGGCCAAGGACGGCTGGATTCAGAACCTGCAGGCGCTGATGGAGAGATTCTTCAGGAGCGAG 
               
               
                   
               
               
                 TCCCGAGGCGCCGTGCGCATCAAGGTGCTGGACGTGCTGTCCTTTGTGCTGCTCATCAACAGGCAGTTCTAT 
               
               
                   
               
               
                 GAGGAGGAGCTGATTAACTCAGTGGTCATCTCGCAGCTCTCCCACATCCCCGAGGATAAAGACCACCAGGTC 
               
               
                   
               
               
                 CGAAAGCTGGCCACCCAGTTGCTGGTGGACCTGGCAGAGGGCTGCCACACACACCACTTCAACAGCCTGCTG 
               
               
                   
               
               
                 GACATCATCGAGAAGGTGATGGCCCGCTCCCTCTCCCCACCCCCGGAGCTGGAAGAAAGGGATGTGGCCGCA 
               
               
                   
               
               
                 TACTCGGCCTCCTTGGAGGATGTGAAGACAGCCGTCCTGGGGCTTCTGGTCATCCTTCAGACCAAGCTGTAC 
               
               
                   
               
               
                 ACCCTGCCTGCAAGCCACGCCACGCGTGTGTATGAGATGCTGGTCAGCCACATTCAGCTCCACTACAAGCAC 
               
               
                   
               
               
                 AGCTACACCCTGCCAATCGCGAGCAGCATCCGGCTGCAGGCC   TTTGACTTCCTGTTGCTGCT   GCGGGCCGAC 
               
               
                   
               
               
                 TCACTGCACCGCCTGGGCCTGCCCAACAAGGATGGAGTCGTGCGGTTCAGCCCCTACTGCGTCTGCGACTAC 
               
               
                   
               
               
                 ATGGAGCCAGAGAGAGGCTCTGAGAAGAAGACCAGCGGCCCCCTTTCTCCTCCCACAGGGCCTCCTGGCCCG 
               
               
                   
               
               
                 GCGCCTGCAGGCCCCGCCGTGCGGCTGGGGTCCGTGCCCTACTCCCTGCTCTTCCGCGTCCTGCTGCAGTGC 
               
               
                   
               
               
                 TTGAAGCAG GAGTCTGACTGGAAGGTGCTGAAGCTGGTTCTGGGCAGGCTGCCTGAGTCCCTGCGCTATAAA   
               
               
                   
               
               
                   GTGCTCATCTTTACTTCCCCTTGCAGTGTGGACCAGCTGTGCTCTGCTCTCTGCTCCATG CTTTCAGGCCCA 
               
               
                   
               
               
                 AAGACACTGGAGCGGCTCCGAGGCGCCCCAGAAGGCTTCTCCAGAACTGACTTGCACCTGGCCGTGGTTCCA 
               
               
                   
               
               
                 GTGCTGACAGCATTAATCTCTTACCATAACTACCTGGACAAAACCAAACAGCGCGAGATGGTCTACTGCCTG 
               
               
                   
               
               
                 GAGCAGGGCCTCATCCACCGCTGTGCCAGCCAGTGCGTCGTGGCCTTGTCCATCTGCAGCGTGGAGATGCCT 
               
               
                   
               
               
                 GACATCATCATCAAGGCGCTGCCTGTTCTGGTGGTGAAGCTCACGCACATCTCAGCCACAGCCAGCATGGCC 
               
               
                   
               
               
                 GTCCCACTGCTGGAGTTCCTGTCCACTCTGGCCAGGCTGCCGCACCTCTACAGGAACTTTGCCGCGGAGCAG 
               
               
                   
               
               
                 TATGCCAGTGTGTTCGCCATCTCCCTGCCGTACACCAACCCCTCCAAGTTTAATCAGTACATCGTGTGTCTG 
               
               
                   
               
               
                 GCCCATCACGTCATAGCCATGTGGTTCATCAGGTGCCGCCTGCCCTTCCGGAAGGATTTTGTCCCTTTCATC 
               
               
                   
               
               
                 ACTAAGGGCCTGCGGTCCAATGTCCTCTTGTCTTTTGATGACACCCCCGAGAAGGACAGCTTCAGGGCCCGG 
               
               
                   
               
               
                 AGTACTAGTCTCAACGAGAGACCCAAGAGTCTGAGGATAGCCAGACCCCCCAAACAAGGCTTGAATAACTCT 
               
               
                   
               
               
                 CCACCCGTGAAAGAATTCAAGGAGAGCTCTGCAGCCGAGGCCTTCCGGTGCCGCAGCATCAGTGTGTCTGAA 
               
               
                   
               
               
                 CATGTGGTCCGCAGCAGGATACAGACGTCCCTCACCAGTGCCAGCTTGGGGTCTGCAGATGAGAACTCCGTG 
               
               
                   
               
               
                 GCCCAGGCTGACGATAGCCTGAAAAACCTCCACCTGGAGCTCACGGAAACCTGTCTGGACATGATGGCTCGA 
               
               
                   
               
               
                 TACGTCTTCTCCAACTTCACGGCTGTCCCGAAGAGGTCTCCTGTGGGCGAGTTCCTCCTAGCGGGTGGCAGG 
               
               
                   
               
               
                 ACCAAAACCTGGCTGGTTGGGAACAAGCTTGTCACTGTGACGACAAGCGTGGGAACCGGGACCCGGTCGTTA 
               
               
                   
               
               
                 CTAGGCCTGGACTCGGGGGAGCTGCAGTCCGGCCCGGAGTCGAGCTCCAGCCCCGGGGTGCATGTGAGACAG 
               
               
                   
               
               
                 ACCAAGGAGGCGCCGGCCAAGCTGGAGTCCCAGGCTGGGCAGCAGGTGTCCCGTGGGGCCCGGGATCGGGTC 
               
               
                   
               
               
                 CGTTCCATGTCGGGGGGCCATGGTCTTCGAGTTGGCGCCCTGGACGTGCCGGCCTCCCAGTTCCTGGGCAGT 
               
               
                   
               
               
                 GCCACTTCTCCAGGACCACGGACTGCACCAGCCGCGAAACCTGAGAAGGCCTCAGCTGGCACCCGGGTTCCT 
               
               
                   
               
               
                 GTGCAGGAGAAGACGAACCTGGCGGCCTATGTGCCCCTGCTGACCCAGGGCTGGGCGGAGATCCTGGTCCGG 
               
               
                   
               
               
                 AGGCCCACAGGGAACACCAGCTGGCTGATGAGCCTGGAGAACCCGCTCAGCCCTTTCTCCTCGGACATCAAC 
               
               
                   
               
               
                 AACATGCCCCTGCAGGAGCTGTCTAACGCCCTCATGGCGGCTGAGCGCTTCAAGGAGCACCGGGACACAGCC 
               
               
                   
               
               
                 CTGTACAAGTCACTGTCGGTGCCGGCAGCCAGCACGGCCAAACCCCCTCCTCTGCCTCGCTCCAACACAGTG 
               
               
                   
               
               
                 GCCTCTTTCTCCTCCCTGTACCAGTCCAGCTGCCAAGGACAGCTGCACAGGAGCGTTTCCTGGGCAGACTCC 
               
               
                   
               
               
                 GCCGTGGTCATGGAGGAGGGAAGTCCGGGCGAGGTTCCTGTGCTGGTGGAGCCCCCAGGGTTGGAGGACGTT 
               
               
                   
               
               
                 GAGGCAGCGCTAGGCATGGACAGGCGCACGGATGCCTACAGCAGGTCGTCCTCAGTCTCCAGCCAGGAGGAG 
               
               
                   
               
               
                 AAGTCGCTCCACGCGGAGGAGCTGGTTGGCAGGGGCATCCCCATCGAGCGAGTCGTCTCCTCGGAGGGTGGC 
               
               
                   
               
               
                 CGGCCCTCTGTGGACCTCTCCTTCCAGCCCTCGCAGCCCCTGAGCAAGTCCAGCTCCTCTCCCGAGCTGCAG 
               
               
                   
               
               
                 ACTCTGCAGGACATCCTCGGGGACCCTGGGGACAAGGCCGACGTGGGCCGGCTGAGCCCTGAGGTTAAGGCC 
               
               
                   
               
               
                 CGGTCACAGTCAGGGACCCTGGACGGGGAAAGTGCTGCCTGGTCGGCCTCGGGCGAAGACAGTCGGGGCCAG 
               
               
                   
               
               
                 CCCGAGGGTCCCTTGCCTTCCAGCTCCCCCCGCTCGCCCAGTGGCCTCCGGCCCCGAGGTTACACCATCTCC 
               
               
                   
               
               
                 GACTCGGCCCCATCACGCAGGGGCAAGAGAGTAGAGAGGGACGCCTTAAAGAGCAGAGCCACAGCCTCCAAT 
               
               
                   
               
               
                 GCAGAGAAAGTGCCAGGCATCAACCCCAGTTTCGTGTTCCTGCAGCTCTACCATTCCCCCTTCTTTGGCGAC 
               
               
                   
               
               
                 GAGTCAAACAAGCCAATCCTGCTGCCCAATGAGTCACAGTCCTTTGAGCGGTCGGTGCAGCTCCTCGACCAG 
               
               
                   
               
               
                 ATCCCATCATACGACACCCACAAGATCGCCGTCCTGTATGTTGGAGAAGGCCAGAGCAACAGCGAGCTCGCC 
               
               
                   
               
               
                 ATCCTGTCCAATGAGCATGGCTCCTACAGGTACACGGAGTTCCTGACGGGCCTGGGCCGGCTCATCGAGCTG 
               
               
                   
               
               
                 AAGGACTGCCAGCCGGACAAGGTGTACCTGGGAGGCCTGGACGTGTGTGGTGAGGACGGCCAGTTCACCTAC 
               
               
                   
               
               
                 TGCTGGCACGATGACATCATGCAAGCCGTCTTCCACATCGCCACCCTGATGCCCACCAAGGACGTGGACAAG 
               
               
                   
               
               
                 CACCGCTGCGACAAGAAGCGCCACCTGGGCAACGACTTTGTGTCCATTGTCTACAATGACTCCGGTGAGGAC 
               
               
                   
               
               
                 TTCAAGCTTGGCACCATCAAGGGCCAGTTCAACTTTGTCCACGTGATCGTCACCCCGCTGGACTACGAGTGC 
               
               
                   
               
               
                 AACCTGGTGTCCCTGCAGTGCAGGAAAGACATGGAGGGCCTTGTGGACACCAGCGTGGCCAAGATCGTGTCT 
               
               
                   
               
               
                 GACCGCAACCTGCCCTTCGTGGCCCGCCAGATGGCCCTGCACGCAAATATGGCCTCACAGGTGCATCATAGC 
               
               
                   
               
               
                 CGCTCCAACCCCACCGATATCTACCCCTCCAAGTGGATTGCCCGGCTCCGCCACATCAAGCGGCTCCGCCAG 
               
               
                   
               
               
                 CGGATCTGCGAGGAAGCCGCCTACTCCAACCCCAGCCTACCTCTGGTGCACCCTCCGTCCCATAGCAAAGCC 
               
               
                   
               
               
                 CCTGCACAGACTCCAGCCGAGCCCACACCTGGCTATGAGGTGGGCCAGCGGAAGCGCCTCATCTCCTCGGTG 
               
               
                   
               
               
                 GAGGACTTCACCGAGTTTGTGTGA 
               
               
                   
               
               
                 (Exon 19 is indicated by double underline. Primers useful for detection of exon 19 splicing variants are underlined.) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 14 
               
               
                   
               
               
                 TSC2 variant 1 (lacking exon 19) Nucleotide Sequence 
               
               
                 (5301 nt, SEQ ID No. 34) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGGCCAAACCAACAAGCAAAGATTCAGGCTTGAAGGAGAAGTTTAAGATTCTGTTGGGACTGGGAACACCG 
               
               
                   
               
               
                 AGGCCAAATCCCAGGTCTGCAGAGGGTAAACAGACGGAGTTTATCATCACCGCGGAAATACTGAGAGAACTG 
               
               
                   
               
               
                 AGCATGGAATGTGGCCTCAACAATCGCATCCGGATGATAGGGCAGATTTGTGAAGTCGCAAAAACCAAGAAA 
               
               
                   
               
               
                 TTTGAAGAGCACGCAGTGGAAGCACTCTGGAAGGCGGTCGCGGATCTGTTGCAGCCGGAGCGGCCGCTGGAG 
               
               
                   
               
               
                 GCCCGGCACGCGGTGCTGGCTCTGCTGAAGGCCATCGTGCAGGGGCAGGGCGAGCGTTTGGGGGTCCTCAGA 
               
               
                   
               
               
                 GCCCTCTTCTTTAAGGTCATCAAGGATTACCCTTCCAACGAAGACCTTCACGAAAGGCTGGAGGTTTTCAAG 
               
               
                   
               
               
                 GCCCTCACAGACAATGGGAGACACATCACCTACTTGGAGGAAGAGCTGGCTGACTTTGTCCTGCAGTGGATG 
               
               
                   
               
               
                 GATGTTGGCTTGTCCTCGGAATTCCTTCTGGTGCTGGTGAACTTGGTCAAATTCAATAGCTGTTACCTCGAC 
               
               
                   
               
               
                 GAGTACATCGCAAGGATGGTTCAGATGATCTGTCTGCTGTGCGTCCGGACCGCGTCCTCTGTGGACATAGAG 
               
               
                   
               
               
                 GTCTCCCTGCAGGTGCTGGACGCCGTGGTCTGCTACAACTGCCTGCCGGCTGAGAGCCTCCCGCTGTTCATC 
               
               
                   
               
               
                 GTTACCCTCTGTCGCACCATCAACGTCAAGGAGCTCTGCGAGCCTTGCTGGAAGCTGATGCGGAACCTCCTT 
               
               
                   
               
               
                 GGCACCCACCTGGGCCACAGCGCCATCTACAACATGTGCCACCTCATGGAGGACAGAGCCTACATGGAGGAC 
               
               
                   
               
               
                 GCGCCCCTGCTGAGAGGAGCCGTGTTTTTTGTGGGCATGGCTCTCTGGGGAGCCCACCGGCTCTATTCTCTC 
               
               
                   
               
               
                 AGGAACTCGCCGACATCTGTGTTGCCATCATTTTACCAGGCCATGGCATGTCCGAACGAGGTGGTGTCCTAT 
               
               
                   
               
               
                 GAGATCGTCCTGTCCATCACCAGGCTCATCAAGAAGTATAGGAAGGAGCTCCAGGTGGTGGCGTGGGACATT 
               
               
                   
               
               
                 CTGCTGAACATCATCGAACGGCTCCTTCAGCAGCTCCAGACCTTGGACAGCCCGGAGCTCAGGACCATCGTC 
               
               
                   
               
               
                 CATGACCTGTTGACCACGGTGGAGGAGCTGTGTGACCAGAACGAGTTCCACGGGTCTCAGGAGAGATACTTT 
               
               
                   
               
               
                 GAACTGGTGGAGAGATGTGCGGACCAGAGGCCTGAGTCCTCCCTCCTGAACCTGATCTCCTATAGAGCGCAG 
               
               
                   
               
               
                 TCCATCCACCCGGCCAAGGACGGCTGGATTCAGAACCTGCAGGCGCTGATGGAGAGATTCTTCAGGAGCGAG 
               
               
                   
               
               
                 TCCCGAGGCGCCGTGCGCATCAAGGTGCTGGACGTGCTGTCCTTTGTGCTGCTCATCAACAGGCAGTTCTAT 
               
               
                   
               
               
                 GAGGAGGAGCTGATTAACTCAGTGGTCATCTCGCAGCTCTCCCACATCCCCGAGGATAAAGACCACCAGGTC 
               
               
                   
               
               
                 CGAAAGCTGGCCACCCAGTTGCTGGTGGACCTGGCAGAGGGCTGCCACACACACCACTTCAACAGCCTGCTG 
               
               
                   
               
               
                 GACATCATCGAGAAGGTGATGGCCCGCTCCCTCTCCCCACCCCCGGAGCTGGAAGAAAGGGATGTGGCCGCA 
               
               
                   
               
               
                 TACTCGGCCTCCTTGGAGGATGTGAAGACAGCCGTCCTGGGGCTTCTGGTCATCCTTCAGACCAAGCTGTAC 
               
               
                   
               
               
                 ACCCTGCCTGCAAGCCACGCCACGCGTGTGTATGAGATGCTGGTCAGCCACATTCAGCTCCACTACAAGCAC 
               
               
                   
               
               
                 AGCTACACCCTGCCAATCGCGAGCAGCATCCGGCTGCAGGCCTTTGACTTCCTGTTGCTGCTGCGGGCCGAC 
               
               
                   
               
               
                 TCACTGCACCGCCTGGGCCTGCCCAACAAGGATGGAGTCGTGCGGTTCAGCCCCTACTGCGTCTGCGACTAC 
               
               
                   
               
               
                 ATGGAGCCAGAGAGAGGCTCTGAGAAGAAGACCAGCGGCCCCCTTTCTCCTCCCACAGGGCCTCCTGGCCCG 
               
               
                   
               
               
                 GCGCCTGCAGGCCCCGCCGTGCGGCTGGGGTCCGTGCCCTACTCCCTGCTCTTCCGCGTCCTGCTGCAGTGC 
               
               
                   
               
               
                   TTGAAGCA   GC   TTTCAGGCC CAAAGACACTGGAGCGGCTCCGAGGCGCCCCAGAAGGCTTCTCCAGAACTGAC 
               
               
                   
               
               
                 TTGCACCTGGCCGTGGTTCCAGTGCTGACAGCATTAATCTCTTACCATAACTACCTGGACAAAACCAAACAG 
               
               
                   
               
               
                 CGCGAGATGGTCTACTGCCTGGAGCAGGGCCTCATCCACCGCTGTGCCAGCCAGTGCGTCGTGGCCTTGTCC 
               
               
                   
               
               
                 ATCTGCAGCGTGGAGATGCCTGACATCATCATCAAGGCGCTGCCTGTTCTGGTGGTGAAGCTCACGCACATC 
               
               
                   
               
               
                 TCAGCCACAGCCAGCATGGCCGTCCCACTGCTGGAGTTCCTGTCCACTCTGGCCAGGCTGCCGCACCTCTAC 
               
               
                   
               
               
                 AGGAACTTTGCCGCGGAGCAGTATGCCAGTGTGTTCGCCATCTCCCTGCCGTACACCAACCCCTCCAAGTTT 
               
               
                   
               
               
                 AATCAGTACATCGTGTGTCTGGCCCATCACGTCATAGCCATGTGGTTCATCAGGTGCCGCCTGCCCTTCCGG 
               
               
                   
               
               
                 AAGGATTTTGTCCCTTTCATCACTAAGGGCCTGCGGTCCAATGTCCTCTTGTCTTTTGATGACACCCCCGAG 
               
               
                   
               
               
                 AAGGACAGCTTCAGGGCCCGGAGTACTAGTCTCAACGAGAGACCCAAGAGTCTGAGGATAGCCAGACCCCCC 
               
               
                   
               
               
                 AAACAAGGCTTGAATAACTCTCCACCCGTGAAAGAATTCAAGGAGAGCTCTGCAGCCGAGGCCTTCCGGTGC 
               
               
                   
               
               
                 CGCAGCATCAGTGTGTCTGAACATGTGGTCCGCAGCAGGATACAGACGTCCCTCACCAGTGCCAGCTTGGGG 
               
               
                   
               
               
                 TCTGCAGATGAGAACTCCGTGGCCCAGGCTGACGATAGCCTGAAAAACCTCCACCTGGAGCTCACGGAAACC 
               
               
                   
               
               
                 TGTCTGGACATGATGGCTCGATACGTCTTCTCCAACTTCACGGCTGTCCCGAAGAGGTCTCCTGTGGGCGAG 
               
               
                   
               
               
                 TTCCTCCTAGCGGGTGGCAGGACCAAAACCTGGCTGGTTGGGAACAAGCTTGTCACTGTGACGACAAGCGTG 
               
               
                   
               
               
                 GGAACCGGGACCCGGTCGTTACTAGGCCTGGACTCGGGGGAGCTGCAGTCCGGCCCGGAGTCGAGCTCCAGC 
               
               
                   
               
               
                 CCCGGGGTGCATGTGAGACAGACCAAGGAGGCGCCGGCCAAGCTGGAGTCCCAGGCTGGGCAGCAGGTGTCC 
               
               
                   
               
               
                 CGTGGGGCCCGGGATCGGGTCCGTTCCATGTCGGGGGGCCATGGTCTTCGAGTTGGCGCCCTGGACGTGCCG 
               
               
                   
               
               
                 GCCTCCCAGTTCCTGGGCAGTGCCACTTCTCCAGGACCACGGACTGCACCAGCCGCGAAACCTGAGAAGGCC 
               
               
                   
               
               
                 TCAGCTGGCACCCGGGTTCCTGTGCAGGAGAAGACGAACCTGGCGGCCTATGTGCCCCTGCTGACCCAGGGC 
               
               
                   
               
               
                 TGGGCGGAGATCCTGGTCCGGAGGCCCACAGGGAACACCAGCTGGCTGATGAGCCTGGAGAACCCGCTCAGC 
               
               
                   
               
               
                 CCTTTCTCCTCGGACATCAACAACATGCCCCTGCAGGAGCTGTCTAACGCCCTCATGGCGGCTGAGCGCTTC 
               
               
                   
               
               
                 AAGGAGCACCGGGACACAGCCCTGTACAAGTCACTGTCGGTGCCGGCAGCCAGCACGGCCAAACCCCCTCCT 
               
               
                   
               
               
                 CTGCCTCGCTCCAACACAGTGGCCTCTTTCTCCTCCCTGTACCAGTCCAGCTGCCAAGGACAGCTGCACAGG 
               
               
                   
               
               
                 AGCGTTTCCTGGGCAGACTCCGCCGTGGTCATGGAGGAGGGAAGTCCGGGCGAGGTTCCTGTGCTGGTGGAG 
               
               
                   
               
               
                 CCCCCAGGGTTGGAGGACGTTGAGGCAGCGCTAGGCATGGACAGGCGCACGGATGCCTACAGCAGGTCGTCC 
               
               
                   
               
               
                 TCAGTCTCCAGCCAGGAGGAGAAGTCGCTCCACGCGGAGGAGCTGGTTGGCAGGGGCATCCCCATCGAGCGA 
               
               
                   
               
               
                 GTCGTCTCCTCGGAGGGTGGCCGGCCCTCTGTGGACCTCTCCTTCCAGCCCTCGCAGCCCCTGAGCAAGTCC 
               
               
                   
               
               
                 AGCTCCTCTCCCGAGCTGCAGACTCTGCAGGACATCCTCGGGGACCCTGGGGACAAGGCCGACGTGGGCCGG 
               
               
                   
               
               
                 CTGAGCCCTGAGGTTAAGGCCCGGTCACAGTCAGGGACCCTGGACGGGGAAAGTGCTGCCTGGTCGGCCTCG 
               
               
                   
               
               
                 GGCGAAGACAGTCGGGGCCAGCCCGAGGGTCCCTTGCCTTCCAGCTCCCCCCGCTCGCCCAGTGGCCTCCGG 
               
               
                   
               
               
                 CCCCGAGGTTACACCATCTCCGACTCGGCCCCATCACGCAGGGGCAAGAGAGTAGAGAGGGACGCCTTAAAG 
               
               
                   
               
               
                 AGCAGAGCCACAGCCTCCAATGCAGAGAAAGTGCCAGGCATCAACCCCAGTTTCGTGTTCCTGCAGCTCTAC 
               
               
                   
               
               
                 CATTCCCCCTTCTTTGGCGACGAGTCAAACAAGCCAATCCTGCTGCCCAATGAGTCACAGTCCTTTGAGCGG 
               
               
                   
               
               
                 TCGGTGCAGCTCCTCGACCAGATCCCATCATACGACACCCACAAGATCGCCGTCCTGTATGTTGGAGAAGGC 
               
               
                   
               
               
                 CAGAGCAACAGCGAGCTCGCCATCCTGTCCAATGAGCATGGCTCCTACAGGTACACGGAGTTCCTGACGGGC 
               
               
                   
               
               
                 CTGGGCCGGCTCATCGAGCTGAAGGACTGCCAGCCGGACAAGGTGTACCTGGGAGGCCTGGACGTGTGTGGT 
               
               
                   
               
               
                 GAGGACGGCCAGTTCACCTACTGCTGGCACGATGACATCATGCAAGCCGTCTTCCACATCGCCACCCTGATG 
               
               
                   
               
               
                 CCCACCAAGGACGTGGACAAGCACCGCTGCGACAAGAAGCGCCACCTGGGCAACGACTTTGTGTCCATTGTC 
               
               
                   
               
               
                 TACAATGACTCCGGTGAGGACTTCAAGCTTGGCACCATCAAGGGCCAGTTCAACTTTGTCCACGTGATCGTC 
               
               
                   
               
               
                 ACCCCGCTGGACTACGAGTGCAACCTGGTGTCCCTGCAGTGCAGGAAAGACATGGAGGGCCTTGTGGACACC 
               
               
                   
               
               
                 AGCGTGGCCAAGATCGTGTCTGACCGCAACCTGCCCTTCGTGGCCCGCCAGATGGCCCTGCACGCAAATATG 
               
               
                   
               
               
                 GCCTCACAGGTGCATCATAGCCGCTCCAACCCCACCGATATCTACCCCTCCAAGTGGATTGCCCGGCTCCGC 
               
               
                   
               
               
                 CACATCAAGCGGCTCCGCCAGCGGATCTGCGAGGAAGCCGCCTACTCCAACCCCAGCCTACCTCTGGTGCAC 
               
               
                   
               
               
                 CCTCCGTCCCATAGCAAAGCCCCTGCACAGACTCCAGCCGAGCCCACACCTGGCTATGAGGTGGGCCAGCGG 
               
               
                   
               
               
                 AAGCGCCTCATCTCCTCGGTGGAGGACTTCACCGAGTTTGTGTGA 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
               
             
           
               
                 TABLE 15 
               
               
                   
               
               
                 Primer across the junction between 
               
               
                 TSC2 exon 18 and 20 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Table 23. Primer sequences 
                 
                   CTTGAAGCA 
                   GC 
                   TTTCAGGCC 
                 
               
               
                 across the junction between 
                   
               
               
                 TSC2 exon 18 and 20 
                   
               
               
                 (SEQ ID No. 33) 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
           
               
                 TABLE 16 
               
               
                   
               
               
                 siRNA for selectively knockdown TSC2 full 
               
               
                 length and variant expression 
               
               
                   
               
             
             
               
                 siRNA targeting TSC2 exon 19 
               
               
                 Sense (SEQ ID No. 31) 5′  CUGCGCUAUAAAGUGCUCA dTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 32) 3′ dTdT GACGCGAUAUUUCACG-   
               
               
                   AGU  (5′-P)5′ 
               
               
                   
               
               
                 siRNA targeting the junction between 
               
               
                 TSC2 exon 18 and exon 20 
               
               
                 Sense (SEQ ID No. 35) 5′ GAAGCA GC UUUCAGGCCCAdTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 36) 3′ dTdTCUUCGU CG AAAGUCCG- 
               
               
                 GGU (5′-P)5′ 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 17 
               
               
                   
               
               
                 RASGRP2 (full length) Nucleotide Sequence (1830 nt, SEQ ID No. 37) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGGCAGGCACCCTGGACCTGGACAAGGGCTGCACGGTGGAGGAGCTGCTCCGCGGGTGCATCGAAGCCTTC 
               
               
                   
               
               
                 GATGACTCCGGGAAGGTGCGGGACCCGCAGCTGGTGCGCATGTTCCTCATGATGCACCCCTGGTACATCCCC 
               
               
                   
               
               
                 TCCTCTCAGCTGGCGGCCAAGCTGCTCCACATCTACCAACAATCCCGGAAGGACAACTCCAATTCCCTGCAG 
               
               
                   
               
               
                 GTGAAAACGTGCCACCTGGTCAGGTACTGGATCTCCGCCTTCCCAGCGGAGTTTGACTTGAACCCGGAGTTG 
               
               
                   
               
               
                 GCTGAGCAGATCAAGGAGCTGAAGGCTCTGCTAGACCAAGAAGGGAACCGACGGCACAGCAGCCTAATCGAC 
               
               
                   
               
               
                 ATAGACAGCGTCCCTACCTACAAGTGGAAGCGGCAGGTGACTCAGCGGAACCCTGTGGGACAGAAAAAGCGC 
               
               
                   
               
               
                 AAGATGTCCCTGTTGTTTGACCACCTGGAGCCCATGGAGCTGGCGGAGCATCTCACCTACTTGGAGTATCGC 
               
               
                   
               
               
                 TCCTTCTGCAAGATCCTGTTTCAGGACTATCACAGTTTCGTGACTCATGGCTGCACTGTGGACAACCCCGTC 
               
               
                   
               
               
                 CTGGAGCGGTTCATCTCCCTCTTCAACAGCGTCTCACAGTGGGTGCAGCTCATGATCCTCAGCAAACCCACA 
               
               
                   
               
               
                 GCCCCGCAGCGGGCCCTGGTCATCACACACTTTGTCCACGTGGCGGAGAAGCTGCTACAGCTGCAGAACTTC 
               
               
                   
               
               
                 AACACGCTGATGGCAGTGGTCGGGGGCCTGAGCCACAGCTCCATCTCCCGCCTCAAGGAGACCCACAGCCAC 
               
               
                   
               
               
                 GTTAGCCCTGAGACCATCAAGCTCTGGGAGGGTCTCACGGAACTAGTGACGGCGACAGGCAACTATGGCAAC 
               
               
                   
               
               
                 TACCGGCGTCGGCTGGCAGCCTGTGTGGGCTTCCGCTTCCCGATCCTGGGTGTGCACCTCAAGGACCTGGTG 
               
               
                   
               
               
                 GCCCTGCAGCTGGCACTGCCTGACTGGCTGGACCCAGCCCGGACCCGGCTCAACGGGGCCAAGATGAAGCAG 
               
               
                   
               
               
                 CTCTTTAGCATCCTGGAGGAGCTGGCCATGGTGACCAGCCTGCGGCCACCAGTACAGGCCAACCCCGACCTG 
               
               
                   
               
               
                 CTGAGCCTGC TCACGGTGTCTCTGGATCAGT ATCAGACGGAGGATGAGCTGTACCAGCTGTCCCTGCAGCGG 
               
               
                   
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                 
                   
                             
                     
                         
                         
                     
                   
                 
               
               
                   
               
               
                 GTTTCCTATTTCCTGCGCTCCAGCTCTGTGTTGGGGGGGCGCATGGGCTTCGTACACAACTTCCAGGAGAGC 
               
               
                   
               
               
                 AACTCCTTGCGCCCCGTCGCCTGCCGCCACTGCAAAGCCCTGATCCTGGGCATCTACAAGCAGGGCCTCAAA 
               
               
                   
               
               
                 TGCCGAGCCTGTGGAGTGAACTGCCACAAGCAGTGCAAGGATCGCCTGTCAGTTGAGTGTCGGCGCAGGGCC 
               
               
                   
               
               
                 CAGAGTGTGAGCCTGGAGGGGTCTGCACCCTCACCCTCACCCATGCACAGCCACCATCACCGCGCCTTCAGC 
               
               
                   
               
               
                 TTCTCTCTGCCCCGCCCTGGCAGGCGAGGCTCCAGGCCTCCAGAGATCCGTGAGGAGGAGGTACAGACGGTG 
               
               
                   
               
               
                 GAGGATGGGGTGTTTGACATCCACTTGTAA 
               
               
                   
               
               
                 (Exon 10 is indicated by double underline. Exon 11 is indicated by wave underline.) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 18 
               
               
                   
               
               
                 RASGRP2 variant 1 (lacking exon 10) 
               
               
                 Nucleotide Sequence (1707 nt, 
               
               
                 SEQ ID No. 45) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGGCAGGCACCCTGGACCTGGACAAGGGCTGCACGGTGGAGGAGCTGCT 
               
               
                   
               
               
                 CCGCGGGTGCATCGAAGCCTTCGATGACTCCGGGAAGGTGCGGGACCCGC 
               
               
                   
               
               
                 AGCTGGTGCGCATGTTCCTCATGATGCACCCCTGGTACATCCCCTCCTCT 
               
               
                   
               
               
                 CAGCTGGCGGCCAAGCTGCTCCACATCTACCAACAATCCCGGAAGGACAA 
               
               
                   
               
               
                 CTCCAATTCCCTGCAGGTGAAAACGTGCCACCTGGTCAGGTACTGGATCT 
               
               
                   
               
               
                 CCGCCTTCCCAGCGGAGTTTGACTTGAACCCGGAGTTGGCTGAGCAGATC 
               
               
                   
               
               
                 AAGGAGCTGAAGGCTCTGCTAGACCAAGAAGGGAACCGACGGCACAGCAG 
               
               
                   
               
               
                 CCTAATCGACATAGACAGCGTCCCTACCTACAAGTGGAAGCGGCAGGTGA 
               
               
                   
               
               
                 CTCAGCGGAACCCTGTGGGACAGAAAAAGCGCAAGATGTCCCTGTTGTTT 
               
               
                   
               
               
                 GACCACCTGGAGCCCATGGAGCTGGCGGAGCATCTCACCTACTTGGAGTA 
               
               
                   
               
               
                 TCGCTCCTTCTGCAAGATCCTGTTTCAGGACTATCACAGTTTCGTGACTC 
               
               
                   
               
               
                 ATGGCTGCACTGTGGACAACCCCGTCCTGGAGCGGTTCATCTCCCTCTTC 
               
               
                   
               
               
                 AACAGCGTCTCACAGTGGGTGCAGCTCATGATCCTCAGCAAACCCACAGC 
               
               
                   
               
               
                 CCCGCAGCGGGCCCTGGTCATCACACACTTTGTCCACGTGGCGGAGAAGC 
               
               
                   
               
               
                 TGCTACAGCTGCAGAACTTCAACACGCTGATGGCAGTGGTCGGGGGCCTG 
               
               
                   
               
               
                 AGCCACAGCTCCATCTCCCGCCTCAAGGAGACCCACAGCCACGTTAGCCC 
               
               
                   
               
               
                 TGAGACCATCAAGCTCTGGGAGGGTCTCACGGAACTAGTGACGGCGACAG 
               
               
                   
               
               
                 GCAACTATGGCAACTACCGGCGTCGGCTGGCAGCCTGTGTGGGCTTCCGC 
               
               
                   
               
               
                 TTCCCGATCCTGGGTGTGCACCTCAAGGACCTGGTGGCCCTGCAGCTGGC 
               
               
                   
               
               
                 ACTGCCTGACTGGCTGGACCCAGCCCGGACCCGGCTCAACGGGGCCAAGA 
               
               
                   
               
               
                 TGAAGCAGCTCTTTAGCATCCTGGAGGAGCTGGCCATGGTGACCAGCCTG 
               
               
                   
               
               
                 CGGCCACCAGTACAGGCCAACCCCGACCTGCTGAGCCTGCTCACGGTGTC 
               
               
                   
               
               
                 TCTGGATCAGTATCAGACGGAGGATGAGCTGTACCAGCTGTCCCTGCAGC 
               
               
                   
               
               
                 GGGAGCCGCGCTC   CAAGTCCTC       GT       CTGTGTTCC   GGAACTTTGACGTCGAT 
               
               
                   
               
               
                 GGGGATGGCCACATCTCACAGGAAGAATTCCAGATCATCCGTGGGAACTT 
               
               
                   
               
               
                 CCCTTACCTCAGCGCCTTTGGGGACCTCGACCAGAACCAGGATGGCTGCA 
               
               
                   
               
               
                 TCAGCAGGGAGGAGATGGTTTCCTATTTCCTGCGCTCCAGCTCTGTGTTG 
               
               
                   
               
               
                 GGGGGGCGCATGGGCTTCGTACACAACTTCCAGGAGAGCAACTCCTTGCG 
               
               
                   
               
               
                 CCCCGTCGCCTGCCGCCACTGCAAAGCCCTGATCCTGGGCATCTACAAGC 
               
               
                   
               
               
                 AGGGCCTCAAATGCCGAGCCTGTGGAGTGAACTGCCACAAGCAGTGCAAG 
               
               
                   
               
               
                 GATCGCCTGTCAGTTGAGTGTCGGCGCAGGGCCCAGAGTGTGAGCCTGGA 
               
               
                   
               
               
                 GGGGTCTGCACCCTCACCCTCACCCATGCACAGCCACCATCACCGCGCCT 
               
               
                   
               
               
                 TCAGCTTCTCTCTGCCCCGCCCTGGCAGGCGAGGCTCCAGGCCTCCAGAG 
               
               
                   
               
               
                 ATCCGTGAGGAGGAGGTACAGACGGTGGAGGATGGGGTGTTTGACATCCA 
               
               
                   
               
               
                 CTTGTAA 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 19 
               
               
                   
               
               
                 RASGRP2 variant 2 (lacking exon 11) 
               
               
                 Nucleotide Sequence (1714 nt, 
               
               
                 SEQ ID No. 49) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGGCAGGCACCCTGGACCTGGACAAGGGCTGCACGGTGGAGGAGCTGCT 
               
               
                   
               
               
                 CCGCGGGTGCATCGAAGCCTTCGATGACTCCGGGAAGGTGCGGGACCCGC 
               
               
                   
               
               
                 AGCTGGTGCGCATGTTCCTCATGATGCACCCCTGGTACATCCCCTCCTCT 
               
               
                   
               
               
                 CAGCTGGCGGCCAAGCTGCTCCACATCTACCAACAATCCCGGAAGGACAA 
               
               
                   
               
               
                 CTCCAATTCCCTGCAGGTGAAAACGTGCCACCTGGTCAGGTACTGGATCT 
               
               
                   
               
               
                 CCGCCTTCCCAGCGGAGTTTGACTTGAACCCGGAGTTGGCTGAGCAGATC 
               
               
                   
               
               
                 AAGGAGCTGAAGGCTCTGCTAGACCAAGAAGGGAACCGACGGCACAGCAG 
               
               
                   
               
               
                 CCTAATCGACATAGACAGCGTCCCTACCTACAAGTGGAAGCGGCAGGTGA 
               
               
                   
               
               
                 CTCAGCGGAACCCTGTGGGACAGAAAAAGCGCAAGATGTCCCTGTTGTTT 
               
               
                   
               
               
                 GACCACCTGGAGCCCATGGAGCTGGCGGAGCATCTCACCTACTTGGAGTA 
               
               
                   
               
               
                 TCGCTCCTTCTGCAAGATCCTGTTTCAGGACTATCACAGTTTCGTGACTC 
               
               
                   
               
               
                 ATGGCTGCACTGTGGACAACCCCGTCCTGGAGCGGTTCATCTCCCTCTTC 
               
               
                   
               
               
                 AACAGCGTCTCACAGTGGGTGCAGCTCATGATCCTCAGCAAACCCACAGC 
               
               
                   
               
               
                 CCCGCAGCGGGCCCTGGTCATCACACACTTTGTCCACGTGGCGGAGAAGC 
               
               
                   
               
               
                 TGCTACAGCTGCAGAACTTCAACACGCTGATGGCAGTGGTCGGGGGCCTG 
               
               
                   
               
               
                 AGCCACAGCTCCATCTCCCGCCTCAAGGAGACCCACAGCCACGTTAGCCC 
               
               
                   
               
               
                 TGAGACCATCAAGCTCTGGGAGGGTCTCACGGAACTAGTGACGGCGACAG 
               
               
                   
               
               
                 GCAACTATGGCAACTACCGGCGTCGGCTGGCAGCCTGTGTGGGCTTCCGC 
               
               
                   
               
               
                 TTCCCGATCCTGGGTGTGCACCTCAAGGACCTGGTGGCCCTGCAGCTGGC 
               
               
                   
               
               
                 ACTGCCTGACTGGCTGGACCCAGCCCGGACCCGGCTCAACGGGGCCAAGA 
               
               
                   
               
               
                 TGAAGCAGCTCTTTAGCATCCTGGAGGAGCTGGCCATGGTGACCAGCCTG 
               
               
                   
               
               
                 CGGCCACCAGTACAGGCCAACCCCGACCTGCTGAGCCTGCTCACGGTGTC 
               
               
                   
               
               
                 TCTGGATCAGTATCAGACGGAGGATGAGCTGTACCAGCTGTCCCTGCAGC 
               
               
                   
               
               
                 GGGAGCCGCGCTCCAAGTCCTCGCCAACCAGCCCCACGAGTTGCACCCCA 
               
               
                   
               
               
                 CCACCCCGGCCCCCGGTACTGGAGGAGTGGACCTCGGCTGCCAAACCCAA 
               
               
                   
               
               
                 GCTGGATCAGGCCCTCGTGGTGGAGCACATCGAGAA GATGGTGGA   GG   GAT   
               
               
                   
               
               
                   GGCTGC ATCAGCAGGGAGGAGATGGTTTCCTATTTCCTGCGCTCCAGCTC 
               
               
                   
               
               
                 TGTGTTGGGGGGGCGCATGGGCTTCGTACACAACTTCCAGGAGAGCAACT 
               
               
                   
               
               
                 CCTTGCGCCCCGTCGCCTGCCGCCACTGCAAAGCCCTGATCCTGGGCATC 
               
               
                   
               
               
                 TACAAGCAGGGCCTCAAATGCCGAGCCTGTGGAGTGAACTGCCACAAGCA 
               
               
                   
               
               
                 GTGCAAGGATCGCCTGTCAGTTGAGTGTCGGCGCAGGGCCCAGAGTGTGA 
               
               
                   
               
               
                 GCCTGGAGGGGTCTGCACCCTCACCCTCACCCATGCACAGCCACCATCAC 
               
               
                   
               
               
                 CGCGCCTTCAGCTTCTCTCTGCCCCGCCCTGGCAGGCGAGGCTCCAGGCC 
               
               
                   
               
               
                 TCCAGAGATCCGTGAGGAGGAGGTACAGACGGTGGAGGATGGGGTGTTTG 
               
               
                   
               
               
                 ACATCCACTTGTAA 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
           
               
                 TABLE 20 
               
               
                   
               
               
                 Primer across the junction between 
               
               
                 RASGRP2 variants 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Primer across junction  
                 
                   
                     CAAGTCCTC 
                   
                   
                     GT 
                   
                   
                     CTGTGTTCC 
                   
                 
               
               
                   
                 between RASGRP2 exon 
                   
               
               
                   
                 9 and exon 11 
                   
               
               
                   
                 (SEQ ID No. 44) 
                   
               
               
                   
                   
               
               
                   
                 Primer across junction  
                 
                   GATGGTGGA 
                   GG 
                   GATGGCTGC 
                 
               
               
                   
                 between RASGRP2 exon 
                   
               
               
                   
                 10 and exon 12 
                   
               
               
                   
                 (SEQ ID No. 48) 
               
               
                   
                   
               
               
                   
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
           
               
                 TABLE 21 
               
               
                   
               
               
                 siRNA for selectively knockdown 
               
               
                 RASGRP2 full length and variants 
               
               
                 expression 
               
               
                   
               
             
             
               
                 siRNA targeting RASGRP2 exon 10 
               
               
                 Sense (SEQ ID No. 40): 5′ GUGGAGCACAUCGAGAAGAdTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 41): 3′ dTdTCACCUCGUGUAGCUC- 
               
               
                 UUCU (5′-P)5′ 
               
               
                   
               
               
                 siRNA targeting RASGRP2 exon 11 
               
               
                 Sense (SEQ ID No. 42): 5′ CCACAUCUCACAGGAAGAAdTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 43): 3′ dTdTGGUGUAGAGUGUCCU- 
               
               
                 UCUU (5′-P)5 
               
               
                   
               
               
                 siRNA targeting junction between 
               
               
                 RASGRP2 exon 9 and 11: 
               
               
                 Sense (SEQ ID No. 46): 5′ CCUC GU CUGUGUUCCGGAAdTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 47): 3′ dTdTGGAG CA GACACAAGG- 
               
               
                 CCUU (5′-P)5′ 
               
               
                   
               
               
                 siRNA targeting junction between 
               
               
                 RASGRP2 exon 10 and 12 
               
               
                 Sense (SEQ ID No. 50): 5′ GGUGGA GG GAUGGCUGCAUdTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 51): 3′ dTdTCCACCU CC CUACCGA- 
               
               
                 CGUA (5′-P)5′ 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 22 
               
               
                   
               
               
                 ITGA4 (full length) Nucleotide Sequence 
               
               
                 (3099 nt, SEQ ID No. 52) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGGCTTGGGAAGCGAGGCGCGAACCCGGCCCCCGAAGGGCCGCCGTCCG 
               
               
                   
               
               
                 GGAGACGGTGATGCTGTTGCTGTGCCTGGGGGTCCCGACCGGCCGCCCCT 
               
               
                   
               
               
                 ACAACGTGGACACTGAGAGCGCGCTGCTTTACCAGGGCCCCCACAACACG 
               
               
                   
               
               
                 CTGTTCGGCTACTCGGTCGTGCTGCACAGCCACGGGGCGAACCGATGGCT 
               
               
                   
               
               
                 CCTAGTGGGTGCGCCCACTGCCAACTGGCTCGCCAACGCTTCAGTGATCA 
               
               
                   
               
               
                 ATCCCGGGGCGATTTACAGATGCAGGATCGGAAAGAATCCCGGCCAGACG 
               
               
                   
               
               
                 TGCGAACAGCTCCAGCTGGGTAGCCCTAATGGAGAACCTTGTGGAAAGAC 
               
               
                   
               
               
                 TTGTTTGGAAGAGAGAGACAATCAGTGGTTGGGGGTCACACTTTCCAGAC 
               
               
                   
               
               
                 AGCCAGGAGAAAATGGATCCATCGTGACTTGTGGGCATAGATGGAAAAAT 
               
               
                   
               
               
                 ATATTTTACATAAAGAATGAAAATAAGCTCCCCACTGGTGGTTGCTATGG 
               
               
                   
               
               
                 AGTGCCCCCTGATTTACGAACAGAACTGAGTAAAAGAATAGCTCCGTGTT 
               
               
                   
               
               
                 ATCAAGATTATGTGAAAAAATTTGGAGAAAATTTTGCATCATGTCAAGCT 
               
               
                   
               
               
                 GGAATATCCAGTTTTTACACAAAGGATTTAATTGTGATGGGGGCCCCAGG 
               
               
                   
               
               
                 ATCATCTTACTGGACTGGCTCTCTTTTTGTCTACAATATAACTACAAATA 
               
               
                   
               
               
                 AATACAAGGCTTTTTTAGACAAACAAAATCAAGTAAAATTTGGAAGTTAT 
               
               
                   
               
               
                 TTAGGATATTCAGTCGGAGCTGGTCATTTTCGGAGCCAGCATACTACCGA 
               
               
                   
               
               
                 AGTAGTCGGAGGAGCTCCTCAACATGAGCAGATTGGTAAGGCATATATAT 
               
               
                   
               
               
                 TCAGCATTGATGAAAAAGAACTAAATATCTTACATGAAATGAAAGGTAAA 
               
               
                   
               
               
                 AAGCTTGGATCGTACTTTGGAGCTTCTGTCTGTGCTGTGGACCTCAATGC 
               
               
                   
               
               
                 AGATGGCTTCTCAGATCTGCTCGTGGGAGCACCCATGCAGAGCACCATCA 
               
               
                   
               
               
                 GAGAGGAAGGAAGAGTGTTTGTGTACATCAACTCTGGCTCGGGAGCAGTA 
               
               
                   
               
               
                 ATGAATGCAATGGAAACAAACCTCGTTGGAAGTGACAAATATGCTGCAAG 
               
               
                   
               
               
                 ATTTGGGGAATCTATAGTTAATCTTGGCGACATTGACAATGATGGCTTTG 
               
               
                   
               
               
                 AAGATGTTGCTATCGGAGCTCCACAAGAAGATGACTTGCAAGGTGCTATT 
               
               
                   
               
               
                 TATATTTACAATGGCCGTGCAGATGGGATCTCGTCAACCTTCTCACAGAG 
               
               
                   
               
               
                 AATTGAAGGACTTCAGATCAGCAAATCGTTAAGTATGTTTGGACAGTCTA 
               
               
                   
               
               
                 TATCAGGACAAATTGATGCAGATAATAATGGCTATGTAGATGTAGCAGTT 
               
               
                   
               
               
                 GGTGCTTTTCGGTCTGATTCTGCTGTCTTGCTAAGGACAAGACCTGTAGT 
               
               
                   
               
               
                 AATTGTTGACGCTTCTTTAAGCCACCCTGAGTCAGTAAATAGAACGAAAT 
               
               
                   
               
               
                 TTGACTGTGTTGAAAATGGATGGCCTTCTGTGTGCATAGATCTAACACTT 
               
               
                   
               
               
                 TGTTTCTCATATAAGGGCAAGGAAGTTCCAGGTTACATTGTTTTGTTTTA 
               
               
                   
               
               
                 TAACATGAGTTTGGATGTGAACAGAAAGGCAGAGTCTCCACCAAGATTCT 
               
               
                   
               
               
                 ATTTCTCTTCTAATGGAACTTCTGACGTGATTACAGGAAGCATACAGGTG 
               
               
                   
               
               
                 TCCAGCAGAGAAGCTAACTGTAGAACACATCAAGCATTTATGCGGAAAGA 
               
               
                   
               
               
                 TGTGCGGGACATCCTCACCCCAATTCAGATTGAAGCTGCTTACCACCTTG 
               
               
                   
               
               
                 GTCCTCATGTCATCAGTAAACGAAGTACAGAGGAATTCCCACCACTTCAG 
               
               
                   
               
               
                 CCAATTCTTCAGCAGAAGAAAGAAAAAGACATAATGAAAAAAACAATAAA 
               
               
                   
               
               
                 CTTTGCAAGGTTTTGTGCCCATGAAAATTGTTCTGCTGATTTACAGGTTT 
               
               
                   
               
               
                 CTGCAAAGATTGGGTTTTTGAA GCCCCATGAAAATAAAACATA TCTTGCT     
               
               
                   
               
               
                 
                     GTTGGGAGTATGAA GACATTGATGTTGAATGTGTCCTTGTTTAATGCTGG 
                 
               
               
                   
               
               
                 
                   AGATGATGCATATGAAACGACTCTACATGTCAAACTACCCGTGGGTCTTT 
                 
               
               
                   
               
               
                   ATTTCATTAAGATTTTAGAGCTG GAAGAGAAGCAAATAAACTGTGAAGTC 
               
               
                   
               
               
                 ACAGATAACTCTGGCGTGGTACAACTTGACTGCAGTATTGGCTATATATA 
               
               
                   
               
               
                 TGTAGATCATCTCTCAAGGATAGATATTAGCTTTCTCCTGGATGTGAGCT 
               
               
                   
               
               
                 CACTCAGCAGAG   CGGAAGAGGACCTCAGTATCA   CAGTGCATGCTACCTGT 
               
               
                   
               
               
                 GAAAATGAAGAGGAAATGGACAATCTAAAGCACAGCAGAGTGACTGTAGC 
               
               
                   
               
               
                 AATACCTTTAAAATATGAGGTTAAGCTGACTGTTCATGGGTTTGTAAACC 
               
               
                   
               
               
                 CAACTTCATTTGTGTATGGATCAAATGATGAAAATGAGCCTGAAACGTGC 
               
               
                   
               
               
                 ATGGTGGAGAAAATGAACTTAACTTTCCATGTTATCAACACTGGCAATAG 
               
               
                   
               
               
                 TATGGCTCCCAATGTTAGTGTGGAAATAATGGTACCAAATTCTTTTAGCC 
               
               
                   
               
               
                 CCCAAACTGATAAGCTGTTCAACATTTTGGATGTCCAGACTACTACTGGA 
               
               
                   
               
               
                 GAATGCCACTTTGAAAATTATCAAAGAGTGTGTGCATTAGAGCAGCAAAA 
               
               
                   
               
               
                 GAGTGCAATGCAGACCTTGAAAGGCATAGTCCGGTTCTTGTCCAAGACTG 
               
               
                   
               
               
                 ATAAGAGGCTATTGTACTGCATAAAAGCTGATCCACATTGTTTAAATTTC 
               
               
                   
               
               
                 TTGTGTAATTTTGGGAAAATGGAAAGTGGAAAAGAAGCCAGTGTTCATAT 
               
               
                   
               
               
                 CCAACTGGAAGGCCGGCCATCCATTTTAGAAATGGATGAGACTTCAGCAC 
               
               
                   
               
               
                 TCAAGTTTGAAATAAGAGCAACAGGTTTTCCAGAGCCAAATCCAAGAGTA 
               
               
                   
               
               
                 ATTGAACTAAACAAGGATGAGAATGTTGCGCATGTTCTACTGGAAGGACT 
               
               
                   
               
               
                 ACATCATCAAAGACCCAAACGTTATTTCACCATAGTGATTATTTCAAGTA 
               
               
                   
               
               
                 GCTTGCTACTTGGACTTATTGTACTTCTATTGATCTCATATGTTATGTGG 
               
               
                   
               
               
                 AAGGCTGGCTTCTTTAAAAGACAATACAAATCTATCCTACAAGAAGAAAA 
               
               
                   
               
               
                 CAGAAGAGACAGTTGGAGTTATATCAACAGTAAAAGCAATGATGATTAA 
               
               
                   
               
               
                 (Exon 23 is indicated by double underline.) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 23 
               
               
                   
               
               
                 ITGA4 variant (lacking exon 23) Nucleotide 
               
               
                 Sequence (2948 nt, SEQ ID No. 58) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGGCTTGGGAAGCGAGGCGCGAACCCGGCCCCCGAAGGGCCGCCGTCCG 
               
               
                   
               
               
                 GGAGACGGTGATGCTGTTGCTGTGCCTGGGGGTCCCGACCGGCCGCCCCT 
               
               
                   
               
               
                 ACAACGTGGACACTGAGAGCGCGCTGCTTTACCAGGGCCCCCACAACACG 
               
               
                   
               
               
                 CTGTTCGGCTACTCGGTCGTGCTGCACAGCCACGGGGCGAACCGATGGCT 
               
               
                   
               
               
                 CCTAGTGGGTGCGCCCACTGCCAACTGGCTCGCCAACGCTTCAGTGATCA 
               
               
                   
               
               
                 ATCCCGGGGCGATTTACAGATGCAGGATCGGAAAGAATCCCGGCCAGACG 
               
               
                   
               
               
                 TGCGAACAGCTCCAGCTGGGTAGCCCTAATGGAGAACCTTGTGGAAAGAC 
               
               
                   
               
               
                 TTGTTTGGAAGAGAGAGACAATCAGTGGTTGGGGGTCACACTTTCCAGAC 
               
               
                   
               
               
                 AGCCAGGAGAAAATGGATCCATCGTGACTTGTGGGCATAGATGGAAAAAT 
               
               
                   
               
               
                 ATATTTTACATAAAGAATGAAAATAAGCTCCCCACTGGTGGTTGCTATGG 
               
               
                   
               
               
                 AGTGCCCCCTGATTTACGAACAGAACTGAGTAAAAGAATAGCTCCGTGTT 
               
               
                   
               
               
                 ATCAAGATTATGTGAAAAAATTTGGAGAAAATTTTGCATCATGTCAAGCT 
               
               
                   
               
               
                 GGAATATCCAGTTTTTACACAAAGGATTTAATTGTGATGGGGGCCCCAGG 
               
               
                   
               
               
                 ATCATCTTACTGGACTGGCTCTCTTTTTGTCTACAATATAACTACAAATA 
               
               
                   
               
               
                 AATACAAGGCTTTTTTAGACAAACAAAATCAAGTAAAATTTGGAAGTTAT 
               
               
                   
               
               
                 TTAGGATATTCAGTCGGAGCTGGTCATTTTCGGAGCCAGCATACTACCGA 
               
               
                   
               
               
                 AGTAGTCGGAGGAGCTCCTCAACATGAGCAGATTGGTAAGGCATATATAT 
               
               
                   
               
               
                 TCAGCATTGATGAAAAAGAACTAAATATCTTACATGAAATGAAAGGTAAA 
               
               
                   
               
               
                 AAGCTTGGATCGTACTTTGGAGCTTCTGTCTGTGCTGTGGACCTCAATGC 
               
               
                   
               
               
                 AGATGGCTTCTCAGATCTGCTCGTGGGAGCACCCATGCAGAGCACCATCA 
               
               
                   
               
               
                 GAGAGGAAGGAAGAGTGTTTGTGTACATCAACTCTGGCTCGGGAGCAGTA 
               
               
                   
               
               
                 ATGAATGCAATGGAAACAAACCTCGTTGGAAGTGACAAATATGCTGCAAG 
               
               
                   
               
               
                 ATTTGGGGAATCTATAGTTAATCTTGGCGACATTGACAATGATGGCTTTG 
               
               
                   
               
               
                 AAGATGTTGCTATCGGAGCTCCACAAGAAGATGACTTGCAAGGTGCTATT 
               
               
                   
               
               
                 TATATTTACAATGGCCGTGCAGATGGGATCTCGTCAACCTTCTCACAGAG 
               
               
                   
               
               
                 AATTGAAGGACTTCAGATCAGCAAATCGTTAAGTATGTTTGGACAGTCTA 
               
               
                   
               
               
                 TATCAGGACAAATTGATGCAGATAATAATGGCTATGTAGATGTAGCAGTT 
               
               
                   
               
               
                 GGTGCTTTTCGGTCTGATTCTGCTGTCTTGCTAAGGACAAGACCTGTAGT 
               
               
                   
               
               
                 AATTGTTGACGCTTCTTTAAGCCACCCTGAGTCAGTAAATAGAACGAAAT 
               
               
                   
               
               
                 TTGACTGTGTTGAAAATGGATGGCCTTCTGTGTGCATAGATCTAACACTT 
               
               
                   
               
               
                 TGTTTCTCATATAAGGGCAAGGAAGTTCCAGGTTACATTGTTTTGTTTTA 
               
               
                   
               
               
                 TAACATGAGTTTGGATGTGAACAGAAAGGCAGAGTCTCCACCAAGATTCT 
               
               
                   
               
               
                 ATTTCTCTTCTAATGGAACTTCTGACGTGATTACAGGAAGCATACAGGTG 
               
               
                   
               
               
                 TCCAGCAGAGAAGCTAACTGTAGAACACATCAAGCATTTATGCGGAAAGA 
               
               
                   
               
               
                 TGTGCGGGACATCCTCACCCCAATTCAGATTGAAGCTGCTTACCACCTTG 
               
               
                   
               
               
                 GTCCTCATGTCATCAGTAAACGAAGTACAGAGGAATTCCCACCACTTCAG 
               
               
                   
               
               
                 CCAATTCTTCAGCAGAAGAAAGAAAAAGACATAATGAAAAAAACAATAAA 
               
               
                   
               
               
                 CTTTGCAAGGTTTTGTGCCCATGAAAATTGTTCTGCTGATTTACAGGTTT 
               
               
                   
               
               
                 CTGCAAAGATT   GGGTTTTTGA       AG       AAGAGAAGC   AAATAAACTGTGAAGTCA 
               
               
                   
               
               
                 CAGATAACTCTGGCGTGGTACAACTTGACTGCAGTATTGGCTATATATAT 
               
               
                   
               
               
                 GTAGATCATCTCTCAAGGATAGATATTAGCTTTCTCCTGGATGTGAGCTC 
               
               
                   
               
               
                 ACTCAGCAGAGCGGAAGAGGACCTCAGTATCACAGTGCATGCTACCTGTG 
               
               
                   
               
               
                 AAAATGAAGAGGAAATGGACAATCTAAAGCACAGCAGAGTGACTGTAGCA 
               
               
                   
               
               
                 ATACCTTTAAAATATGAGGTTAAGCTGACTGTTCATGGGTTTGTAAACCC 
               
               
                   
               
               
                 AACTTCATTTGTGTATGGATCAAATGATGAAAATGAGCCTGAAACGTGCA 
               
               
                   
               
               
                 TGGTGGAGAAAATGAACTTAACTTTCCATGTTATCAACACTGGCAATAGT 
               
               
                   
               
               
                 ATGGCTCCCAATGTTAGTGTGGAAATAATGGTACCAAATTCTTTTAGCCC 
               
               
                   
               
               
                 CCAAACTGATAAGCTGTTCAACATTTTGGATGTCCAGACTACTACTGGAG 
               
               
                   
               
               
                 AATGCCACTTTGAAAATTATCAAAGAGTGTGTGCATTAGAGCAGCAAAAG 
               
               
                   
               
               
                 AGTGCAATGCAGACCTTGAAAGGCATAGTCCGGTTCTTGTCCAAGACTGA 
               
               
                   
               
               
                 TAAGAGGCTATTGTACTGCATAAAAGCTGATCCACATTGTTTAAATTTCT 
               
               
                   
               
               
                 TGTGTAATTTTGGGAAAATGGAAAGTGGAAAAGAAGCCAGTGTTCATATC 
               
               
                   
               
               
                 CAACTGGAAGGCCGGCCATCCATTTTAGAAATGGATGAGACTTCAGCACT 
               
               
                   
               
               
                 CAAGTTTGAAATAAGAGCAACAGGTTTTCCAGAGCCAAATCCAAGAGTAA 
               
               
                   
               
               
                 TTGAACTAAACAAGGATGAGAATGTTGCGCATGTTCTACTGGAAGGACTA 
               
               
                   
               
               
                 CATCATCAAAGACCCAAACGTTATTTCACCATAGTGATTATTTCAAGTAG 
               
               
                   
               
               
                 CTTGCTACTTGGACTTATTGTACTTCTATTGATCTCATATGTTATGTGGA 
               
               
                   
               
               
                 AGGCTGGCTTCTTTAAAAGACAATACAAATCTATCCTACAAGAAGAAAAC 
               
               
                   
               
               
                 AGAAGAGACAGTTGGAGTTATATCAACAGTAAAAGCAATGATGATTAA 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
               
             
           
               
                 TABLE 24 
               
               
                   
               
               
                 Primer across the junction between ITGA4 
               
               
                 exon 22 and 24 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Primer across the junction 
                 
                   
                     GGGTTTTTGA 
                   
                   
                     AG 
                   
                   
                     AAGAGAAGC 
                   
                 
               
               
                 between ITGA4 exon 22 and 
                   
               
               
                 24 (SEQ ID No. 57) 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
           
               
                 TABLE 25 
               
               
                   
               
               
                 siRNA for selectively knockdown ITGA4 
               
               
                 full length and variants expression 
               
               
                   
               
             
             
               
                 siRNA targeting ITGA4 exon 23 
               
               
                 Sense (SEQ ID No. 55) 5′ GGGAGUAUGAAGACAUUGA dTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 56) 3′ dTdTCCCUCAUACUUCUGUA- 
               
               
                 ACU (5′-P)5′ 
               
               
                   
               
               
                 siRNA targeting splice junction between 
               
               
                 ITGA4 exon 22 and exon 24 
               
               
                 Sense (SEQ ID No. 59) 5′ GA AG AAGAGAAGCAAAUAA dTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 60) 3′ dTdTCU UC UUCUCUUCGUUU- 
               
               
                 AUU (5′-P)5′ 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 26 
               
               
                   
               
               
                 MET (Full length)Nucleotide Sequence (4226 nt, SEQ ID No. 62) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGAAGGCCCCCGCTGTGCTTGCACCTGGCATCCTCGTGCTCCTGTTTACCTTGGTGCAGAGGAGCAATGGG 
               
               
                   
               
               
                 AGTGTAAAGAGGCACTAGCAAAGTCCGAGATGAATGTGAATATGAAGTATCAGCTTCCCAACTTCACCGCGG 
               
               
                   
               
               
                 AAACACCCATCCAGAATGTCATTCTACATGAGCATCACATTTTCCTTGGTGCCACTAACTACATTTATGTTT 
               
               
                   
               
               
                 TAAATGAGGAAGACCTTCAGAAGGTTGCTGAGTACAAGACTGGGCCTGTGCTGGAACACCCAGATTGTTTCC 
               
               
                   
               
               
                 CATGTCAGGACTGCAGCAGCAAAGCCAATTTATCAGGAGGTGTTTGGAAAGATAACATCAACATGGCTCTAG 
               
               
                   
               
               
                 TTGTCGACACCTACTATGATGATCAACTCATTAGCTGTGGCAGCGTCAACAGAGGGACCTGCCAGCGACATG 
               
               
                   
               
               
                 TCTTTCCCCACAATCATACTGCTGACATACAGTCGGAGGTTCACTGCATATTCTCCCCACAGATAGAAGAGC 
               
               
                   
               
               
                 CCAGCCAGTGTCCTGACTGTGTGGTGAGCGCCCTGGGAGCCAAAGTCCTTTCATCTGTAAAGGACCGGTTCA 
               
               
                   
               
               
                 TCAACTTCTTTGTAGGCAATACCATAAATTCTTCTTATTTCCCAGATCATCCATTGCATTCGATATCAGTGA 
               
               
                   
               
               
                 GAAGGCTAAAGGAAACGAAAGATGGTTTTATGTTTTTGACGGACCAGTCCTACATTGATGTTTTACCTGAGT 
               
               
                   
               
               
                 TCAGAGATTCTTACCCCATTAAGTATGTCCATGCCTTTGAAAGCAACAATTTTATTTACTTCTTGACGGTCC 
               
               
                   
               
               
                 AAAGGGAAACTCTAGATGCTCAGACTTTTCACACAAGAATAATCAGGTTCTGTTCCATAAACTCTGGATTGC 
               
               
                   
               
               
                 ATTCCTACATGGAAATGCCTCTGGAGTGTATTCTCACAGAAAAGAGAAAAAAGAGATCCACAAAGAAGGAAG 
               
               
                   
               
               
                 TGTTTAATATACTTCAGGCTGCGTATGTCAGCAAGCCTGGGGCCCAGCTTGCTAGACAAATAGGAGCCAGCC 
               
               
                   
               
               
                 TGAATGATGACATTCTTTTCGGGGTGTTCGCACAAAGCAAGCCAGATTCTGCCGAACCAATGGATCGATCTG 
               
               
                   
               
               
                 CCATGTGTGCATTCCCTATCAAATATGTCAACGACTTCTTCAACAAGATCGTCAACAAAAACAATGTGAGAT 
               
               
                   
               
               
                 GTCTCCAGCATTTTTACGGACCCAATCATGAGCACTGCTTTAATAGGACACTTCTGAGAAATTCATCAGGCT 
               
               
                   
               
               
                 GTGAAGCGCGCCGTGATGAATATCGAACAGAGTTTACCACAGCTTTGCAGCGCGTTGACTTATTCATGGGTC 
               
               
                   
               
               
                 AATTCAGCGAAGTCCTCTTAACATCTATATCCACCTTCATTAAAGGAGACCTCACCATAGCTAATCTTGGGA 
               
               
                   
               
               
                 CATCAGAGGGTCGCTTCATGCAGGTTGTGGTTTCTCGATCAGGACCATCAACCCCTCATGTGAATTTTCTCC 
               
               
                   
               
               
                 TGGACTCCCATCCAGTGTCTCCAGAAGTGATTGTGGAGCATACATTAAACCAAAATGGCTACACACTGGTTA 
               
               
                   
               
               
                 TCACTGGGAAGAAGATCACGAAGATCCCATTGAATGGCTTGGGCTGCAGACATTTCCAGTCCTGCAGTCAAT 
               
               
                   
               
               
                 GCCTCTCTGCCCCACCCTTTGTTCAGTGTGGCTGGTGCCACGACAAATGTGTGCGATCGGAGGAATGCCTGA 
               
               
                   
               
               
                 GCGGGACATGGACTCAACAGATCTGTCTGCCTGCAATCTACAAGGTTTTCCCAAATAGTGCACCCCTTGAAG 
               
               
                   
               
               
                 GAGGGACAAGGCTGACCATATGTGGCTGGGACTTTGGATTTCGGAGGAATAATAAATTTGATTTAAAGAAAA 
               
               
                   
               
               
                 CTAGAGTTCTCCTTGGAAATGAGAGCTGCACCTTGACTTTAAGTGAGAGCACGATGAATACATTGAAATGCA 
               
               
                   
               
               
                 CAGTTGGTCCTGCCATGAATAAGCATTTCAATATGTCCATAATTATTTCAAATGGCCACGGGACAACACAAT 
               
               
                   
               
               
                 ACAGTACATTCTCCTATGTGGATCCTGTAATAACAAGTATTTCGCCGAAATACGGTCCTATGGCTGGTGGCA 
               
               
                   
               
               
                 CTTTACTTACTTTAACTGGAAATTACCTAAACAGTGGGAATTCTAGACACATTTCAATTGGTGGAAAAACAT 
               
               
                   
               
               
                 GTACTTTAAAAAGTGTGTCAAACAGTATTCTTGAATGTTATACCCCAGCCCAAACCATTTCAACTGAGTTTG 
               
               
                   
               
               
                 CTGTTAAATTGAAAATTGACTTAGCCAACCGAGAGACAAGCATCTTCAGTTACCGTGAAGATCCCATTGTCT 
               
               
                   
               
               
                 ATGAAATTCATCCAACCAAATCTTTTATTAGTACTTGGTGGAAAGAACCTCTCAACATTGTCAGTTTTCTAT 
               
               
                   
               
               
                 TTTGCTTTGCCAGTGGTGGGAGCACAATAACAGGTGTTGGGAAAAACCTGAATTCAGTTAGTGTCCCGAGAA 
               
               
                   
               
               
                 TGGTCATAAATGTGCATGAAGCAGGAAGGAACTTTACAGTGGCATGTCAACATCGCTCTAATTCAGAGATAA 
               
               
                   
               
               
                 TCTGTTGTACCACTCCTTCCCTGCAACAGCTGAATCTGCAACTCCCCCTGAAAACCAAAGCCTTTTTCATGT 
               
               
                   
               
               
                 TAGATGGGATCCTTTCCAAATACTTTGATCTCATTTATGTACATAATCCTGTGTTTAAGCCTTTTGAAAAGC 
               
               
                   
               
               
                 CAGTGATGATCTCAATGGGCAATGAAAATGTA   CTGGAAATTAA       GG       GAAATG   ATATTGACCCTGAAGCAGTTA 
               
               
                   
               
               
                 AAGGTGAAGTGTTAAAAGTTGGAAATAAGAGCTGTGAGAATATACACTTACATTCTGAAGCCGTTTTATGCA 
               
               
                   
               
               
                 CGGTCCCCAATGACCTGCTGAAATTGAACAGCGAGCTAAATATAGAGTGGAAGCAAGCAATTTCTTCAACCG 
               
               
                   
               
               
                 TCCTTGGAAAAGTAATAGTTCAACCAGATCAGAATTTCACAGGATTGATTGCTGGTGTTGTCTCAATATCAA 
               
               
                   
               
               
                 CAGCACTGTTATTACTACTTGGGTTTTTCCTGTGGCTGAAAAAGAGAAAGCAAATTAAAGATCTGGGCAGTG 
               
               
                   
               
               
                 AATTAGTTCGCTACGATGCAAGAGTACACACTCCTCATTTGGATAGGCTTGTAAGTGCCCGAAGTGTAAGCC 
               
               
                   
               
               
                 CAACTACAGAAATGGTTTCAAATGAATCTGTAGACTACCGAGCTACTTTTCCAGAAGATCAGTTTCCTAATT 
               
               
                   
               
               
                 CATCTCAGAACGGTTCATGCCGACAAGTGCAGTATCCTCTGACAGACATGTCCCCCATCCTAACTAGTGGGG 
               
               
                   
               
               
                 ACTCTGATATATCCAGTCCATTACTGCAAAATACTGTCCACATTGACCTCAGTGCTCTAAATCCAGAGCTGG 
               
               
                   
               
               
                 TCCAGGCAGTGCAGCATGTAGTGATTGGGCCCAGTAGCCTGATTGTGCATTTCAATGAAGTCATAGGAAGAG 
               
               
                   
               
               
                 GGCATTTTGGTTGTGTATATCATGGGACTTTGTTGGACAATGATGGCAAGAAAATTCACTGTGCTGTGAAAT 
               
               
                   
               
               
                 CCTTGAACAGAATCACTGACATAGGAGAAGTTTCCCAATTTCTGACCGAGGGAATCATCATGAAAGATTTTA 
               
               
                   
               
               
                 GTCATCCCAATGTCCTCTCGCTCCTGGGAATCTGCCTGCGAAGTGAAGGGTCTCCGCTGGTGGTCCTACCAT 
               
               
                   
               
               
                 ACATGAAACATGGAGATCTTCGAAATTTCATTCGAAATGAGACTCATAATCCAACTGTAAAAGATCTTATTG 
               
               
                   
               
               
                 GCTTTGGTCTTCAAGTAGCCAAAGGCATGAAATATCTTGCAAGCAAAAAGTTTGTCCACAGAGACTTGGCTG 
               
               
                   
               
               
                 CAAGAAACTGTATGCTGGATGAAAAATTCACAGTCAAGGTTGCTGATTTTGGTCTTGCCAGAGACATGTATG 
               
               
                   
               
               
                 ATAAAGAATACTATAGTGTACACAACAAAACAGGTGCAAAGCTGCCAGTGAAGTGGATGGCTTTGGAAAGTC 
               
               
                   
               
               
                 TGCAAACTCAAAAGTTTACCACCAAGTCAGATGTGTGGTCCTTTGGCGTGCTCCTCTGGGAGCTGATGACAA 
               
               
                   
               
               
                 GAGGAGCCCCACCTTATCCTGACGTAAACACCTTTGATATAACTGTTTACTTGTTGCAAGGGAGAAGACTCC 
               
               
                   
               
               
                 TACAACCCGAATACTGCCCAGACCCCTTATATGAAGTAATGCTAAAATGCTGGCACCCTAAAGCCGAAATGC 
               
               
                   
               
               
                 GCCCATCCTTTTCTGAACTGGTGTCCCGGATATCAGCGATCTTCTCTACTTTCATTGGGGAGCACTATGTCC 
               
               
                   
               
               
                 ATGTGAACGCTACTTATGTGAACGTAAAATGTGTCGCTCCGTATCCTTCTCTGTTGTCATCAGAAGATAACG 
               
               
                   
               
               
                 CTGATGATGAGGTGGACACACGACCAGCCTCCTTCTGGGAGACATCATAG 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction between exon 26 and 28) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 27 
               
               
                   
               
               
                 MET variant (with non-coding exon 27) Nucleotide 
               
               
                 Sequence (4651 nt, SEQ ID No. 65) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGAAGGCCCCCGCTGTGCTTGCACCTGGCATCCTCGTGCTCCTGTTTACCTTGGTGCAGAGGAGCAATGGG 
               
               
                   
               
               
                 AGTGTAAAGAGGCACTAGCAAAGTCCGAGATGAATGTGAATATGAAGTATCAGCTTCCCAACTTCACCGCGG 
               
               
                   
               
               
                 AAACACCCATCCAGAATGTCATTCTACATGAGCATCACATTTTCCTTGGTGCCACTAACTACATTTATGTTT 
               
               
                   
               
               
                 TAAATGAGGAAGACCTTCAGAAGGTTGCTGAGTACAAGACTGGGCCTGTGCTGGAACACCCAGATTGTTTCC 
               
               
                   
               
               
                 CATGTCAGGACTGCAGCAGCAAAGCCAATTTATCAGGAGGTGTTTGGAAAGATAACATCAACATGGCTCTAG 
               
               
                   
               
               
                 TTGTCGACACCTACTATGATGATCAACTCATTAGCTGTGGCAGCGTCAACAGAGGGACCTGCCAGCGACATG 
               
               
                   
               
               
                 TCTTTCCCCACAATCATACTGCTGACATACAGTCGGAGGTTCACTGCATATTCTCCCCACAGATAGAAGAGC 
               
               
                   
               
               
                 CCAGCCAGTGTCCTGACTGTGTGGTGAGCGCCCTGGGAGCCAAAGTCCTTTCATCTGTAAAGGACCGGTTCA 
               
               
                   
               
               
                 TCAACTTCTTTGTAGGCAATACCATAAATTCTTCTTATTTCCCAGATCATCCATTGCATTCGATATCAGTGA 
               
               
                   
               
               
                 GAAGGCTAAAGGAAACGAAAGATGGTTTTATGTTTTTGACGGACCAGTCCTACATTGATGTTTTACCTGAGT 
               
               
                   
               
               
                 TCAGAGATTCTTACCCCATTAAGTATGTCCATGCCTTTGAAAGCAACAATTTTATTTACTTCTTGACGGTCC 
               
               
                   
               
               
                 AAAGGGAAACTCTAGATGCTCAGACTTTTCACACAAGAATAATCAGGTTCTGTTCCATAAACTCTGGATTGC 
               
               
                   
               
               
                 ATTCCTACATGGAAATGCCTCTGGAGTGTATTCTCACAGAAAAGAGAAAAAAGAGATCCACAAAGAAGGAAG 
               
               
                   
               
               
                 TGTTTAATATACTTCAGGCTGCGTATGTCAGCAAGCCTGGGGCCCAGCTTGCTAGACAAATAGGAGCCAGCC 
               
               
                   
               
               
                 TGAATGATGACATTCTTTTCGGGGTGTTCGCACAAAGCAAGCCAGATTCTGCCGAACCAATGGATCGATCTG 
               
               
                   
               
               
                 CCATGTGTGCATTCCCTATCAAATATGTCAACGACTTCTTCAACAAGATCGTCAACAAAAACAATGTGAGAT 
               
               
                   
               
               
                 GTCTCCAGCATTTTTACGGACCCAATCATGAGCACTGCTTTAATAGGACACTTCTGAGAAATTCATCAGGCT 
               
               
                   
               
               
                 GTGAAGCGCGCCGTGATGAATATCGAACAGAGTTTACCACAGCTTTGCAGCGCGTTGACTTATTCATGGGTC 
               
               
                   
               
               
                 AATTCAGCGAAGTCCTCTTAACATCTATATCCACCTTCATTAAAGGAGACCTCACCATAGCTAATCTTGGGA 
               
               
                   
               
               
                 CATCAGAGGGTCGCTTCATGCAGGTTGTGGTTTCTCGATCAGGACCATCAACCCCTCATGTGAATTTTCTCC 
               
               
                   
               
               
                 TGGACTCCCATCCAGTGTCTCCAGAAGTGATTGTGGAGCATACATTAAACCAAAATGGCTACACACTGGTTA 
               
               
                   
               
               
                 TCACTGGGAAGAAGATCACGAAGATCCCATTGAATGGCTTGGGCTGCAGACATTTCCAGTCCTGCAGTCAAT 
               
               
                   
               
               
                 GCCTCTCTGCCCCACCCTTTGTTCAGTGTGGCTGGTGCCACGACAAATGTGTGCGATCGGAGGAATGCCTGA 
               
               
                   
               
               
                 GCGGGACATGGACTCAACAGATCTGTCTGCCTGCAATCTACAAGGTTTTCCCAAATAGTGCACCCCTTGAAG 
               
               
                   
               
               
                 GAGGGACAAGGCTGACCATATGTGGCTGGGACTTTGGATTTCGGAGGAATAATAAATTTGATTTAAAGAAAA 
               
               
                   
               
               
                 CTAGAGTTCTCCTTGGAAATGAGAGCTGCACCTTGACTTTAAGTGAGAGCACGATGAATACATTGAAATGCA 
               
               
                   
               
               
                 CAGTTGGTCCTGCCATGAATAAGCATTTCAATATGTCCATAATTATTTCAAATGGCCACGGGACAACACAAT 
               
               
                   
               
               
                 ACAGTACATTCTCCTATGTGGATCCTGTAATAACAAGTATTTCGCCGAAATACGGTCCTATGGCTGGTGGCA 
               
               
                   
               
               
                 CTTTACTTACTTTAACTGGAAATTACCTAAACAGTGGGAATTCTAGACACATTTCAATTGGTGGAAAAACAT 
               
               
                   
               
               
                 GTACTTTAAAAAGTGTGTCAAACAGTATTCTTGAATGTTATACCCCAGCCCAAACCATTTCAACTGAGTTTG 
               
               
                   
               
               
                 CTGTTAAATTGAAAATTGACTTAGCCAACCGAGAGACAAGCATCTTCAGTTACCGTGAAGATCCCATTGTCT 
               
               
                   
               
               
                 ATGAAATTCATCCAACCAAATCTTTTATTAGTACT   TGGTGGAAAGAACCTCTCAA   CATTGTCAGTTTTCTAT 
               
               
                   
               
               
                 TTTGCTTTGCCAGTGGTGGGAGCACAATAACAGGTGTTGGGAAAAACCTGAATTCAGTTAGTGTCCCGAGAA 
               
               
                   
               
               
                 TGGTCATAAATGTGCATGAAGCAGGAAGGAACTTTACAGTGGCATGTCAACATCGCTCTAATTCAGAGATAA 
               
               
                   
               
               
                 TCTGTTGTACCACTCCTTCCCTGCAACAGCTGAATCTGCAACTCCCCCTGAAAACCAAAGCCTTTTTCATGT 
               
               
                   
               
               
                 TAGATGGGATCCTTTCCAAATACTTTGATCTCATTTATGTACATAATCCTGTGTTTAAGCCTTTTGAAAAGC 
               
               
                   
               
               
                 CAGTGATGATCTCAATGGGCAATGAAAATGTACTGGAAATTAAG gtgggagcagtggcaattcagggag   
               
               
                   
               
               
                 
                   attattttagtatcatggttcaatattttttcatacttcatttttcttatgtatgagaggaaagc 
                 
               
               
                   
               
               
                 
                   aaaggcataagagaatatttgttgtgtcagcaatctaactctttatcaatacgttaagttgatca 
                 
               
               
                   
               
               
                 
                   cattaaaacttctacctotcagccaggcacggtagctcatacctgtaatcccagcactttgggag 
                 
               
               
                   
               
               
                 
                   gccaaggcgggtgaatcacttgagatcaggagttcaagaccagcctggccaaaatggtgaaaccc 
                 
               
               
                   
               
               
                 
                   catctccactaaaaatacaaaaattagctgggcatggtggtgggtgcctgtaatcccagctactc 
                 
               
               
                   
               
               
                 
                   aggaggctgagggacggaggtgacctgagtcctgaaggcgg aggttgcagtgagccaaga tggca 
                 
               
               
                   
               
               
                   ccactgcact GGAAATGATATTGACCCTGAAGCAGTTAAAGGTGAAGTGTTAAAAGTTGGAAATAAGAGC 
               
               
                   
               
               
                 TGTGAGAATATACACTTACATTCTGAAGCCGTTTTATGCACGGTCCCCAATGACCTGCTGAAATTGAACAGC 
               
               
                   
               
               
                 GAGCTAAATATAGAGTGGAAGCAAGCAATTTCTTCAACCGTCCTTGGAAAAGTAATAGTTCAACCAGATCAG 
               
               
                   
               
               
                 AATTTCACAGGATTGATTGCTGGTGTTGTCTCAATATCAACAGCACTGTTATTACTACTTGGGTTTTTCCTG 
               
               
                   
               
               
                 TGGCTGAAAAAGAGAAAGCAAATTAAAGATCTGGGCAGTGAATTAGTTCGCTACGATGCAAGAGTACACACT 
               
               
                   
               
               
                 CCTCATTTGGATAGGCTTGTAAGTGCCCGAAGTGTAAGCCCAACTACAGAAATGGTTTCAAATGAATCTGTA 
               
               
                   
               
               
                 GACTACCGAGCTACTTTTCCAGAAGATCAGTTTCCTAATTCATCTCAGAACGGTTCATGCCGACAAGTGCAG 
               
               
                   
               
               
                 TATCCTCTGACAGACATGTCCCCCATCCTAACTAGTGGGGACTCTGATATATCCAGTCCATTACTGCAAAAT 
               
               
                   
               
               
                 ACTGTCCACATTGACCTCAGTGCTCTAAATCCAGAGCTGGTCCAGGCAGTGCAGCATGTAGTGATTGGGCCC 
               
               
                   
               
               
                 AGTAGCCTGATTGTGCATTTCAATGAAGTCATAGGAAGAGGGCATTTTGGTTGTGTATATCATGGGACTTTG 
               
               
                   
               
               
                 TTGGACAATGATGGCAAGAAAATTCACTGTGCTGTGAAATCCTTGAACAGAATCACTGACATAGGAGAAGTT 
               
               
                   
               
               
                 TCCCAATTTCTGACCGAGGGAATCATCATGAAAGATTTTAGTCATCCCAATGTCCTCTCGCTCCTGGGAATC 
               
               
                   
               
               
                 TGCCTGCGAAGTGAAGGGTCTCCGCTGGTGGTCCTACCATACATGAAACATGGAGATCTTCGAAATTTCATT 
               
               
                   
               
               
                 CGAAATGAGACTCATAATCCAACTGTAAAAGATCTTATTGGCTTTGGTCTTCAAGTAGCCAAAGGCATGAAA 
               
               
                   
               
               
                 TATCTTGCAAGCAAAAAGTTTGTCCACAGAGACTTGGCTGCAAGAAACTGTATGCTGGATGAAAAATTCACA 
               
               
                   
               
               
                 GTCAAGGTTGCTGATTTTGGTCTTGCCAGAGACATGTATGATAAAGAATACTATAGTGTACACAACAAAACA 
               
               
                   
               
               
                 GGTGCAAAGCTGCCAGTGAAGTGGATGGCTTTGGAAAGTCTGCAAACTCAAAAGTTTACCACCAAGTCAGAT 
               
               
                   
               
               
                 GTGTGGTCCTTTGGCGTGCTCCTCTGGGAGCTGATGACAAGAGGAGCCCCACCTTATCCTGACGTAAACACC 
               
               
                   
               
               
                 TTTGATATAACTGTTTACTTGTTGCAAGGGAGAAGACTCCTACAACCCGAATACTGCCCAGACCCCTTATAT 
               
               
                   
               
               
                 GAAGTAATGCTAAAATGCTGGCACCCTAAAGCCGAAATGCGCCCATCCTTTTCTGAACTGGTGTCCCGGATA 
               
               
                   
               
               
                 TCAGCGATCTTCTCTACTTTCATTGGGGAGCACTATGTCCATGTGAACGCTACTTATGTGAACGTAAAATGT 
               
               
                   
               
               
                 GTCGCTCCGTATCCTTCTCTGTTGTCATCAGAAGATAACGCTGATGATGAGGTGGACACACGACCAGCCTCC 
               
               
                   
               
               
                 TTCTGGGAGACATCATAG 
               
               
                   
               
               
                 (Exon 27 is indicated as double underline.) 
               
             
          
         
       
     
     
       
         
               
             
               
               
             
           
               
                 TABLE 28 
               
               
                   
               
               
                 Primer across the junction between MET 
               
               
                 exon 26 and 28 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Primer across the junction 
                 
                   
                     CTGGAAATTAA 
                   
                   
                     GG 
                   
                   
                     GAAATG 
                   
                 
               
               
                 between MET exon 26 and  
                   
               
               
                 28 (SEQ ID No. 61): 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
           
               
                 TABLE 29 
               
               
                   
               
               
                 siRNA for selectively knockdown MET full 
               
               
                 length and variants expression 
               
               
                   
               
             
             
               
                 siRNA targeting splice junction between 
               
               
                 MET exon 26 and exon 28 
               
               
                 Sense (SEQ ID No. 63) 5′ GUACUGGAAAUUAA GG GAAdTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 64) 3′ dTdTCAUGACCUUUAAUU CC - 
               
               
                 CUU (5′-P)5′ 
               
               
                   
               
               
                 siRNA targeting non-coding MET exon 27 
               
               
                 Sense (SEQ ID No. 68) 5′ CAGCAAUCUAACUCUUUAUdTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 69) 3′ dTdTGUCGUUAGAUUGAGAA- 
               
               
                 AUA (5′-P)5′ 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 30 
               
               
                   
               
               
                 NF1 (full length)Nucleotide Sequence (8520 nt, SEQ ID No. 70) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGGCCGCGCACAGGCCGGTGGAATGGGTCCAGGCCGTGGTCAGCCGCTTCGACGAGCAGCTTCCAATAA 
               
               
                   
               
               
                 AAACAGGACAGCAGAACACACATACCAAAGTCAGTACTGAGCACAACAAGGAATGTCTAATCAATATTTC 
               
               
                   
               
               
                 CAAATACAAGTTTTCTTTGGTTATAAGCGGCCTCACTACTATTTTAAAGAATGTTAACAATATGAGAATA 
               
               
                   
               
               
                 TTTGGAGAAGCTGCTGAAAAAAATTTATATCTCTCTCAGTTGATTATATTGGATACACTGGAAAAATGTC 
               
               
                   
               
               
                 TTGCTGGGCAACCAAAGGACACAATGAGATTAGATGAAACGATGCTGGTCAAACAGTTGCTGCCAGAAAT 
               
               
                   
               
               
                 CTGCCATTTTCTTCACACCTGTCGTGAAGGAAACCAGCATGCAGCTGAACTTCGGAATTCTGCCTCTGGG 
               
               
                   
               
               
                 GTTTTATTTTCTCTCAGCTGCAACAACTTCAATGCAGTCTTTAGTCGCATTTCTACCAGGTTACAGGAAT 
               
               
                   
               
               
                 TAACTGTTTGTTCAGAAGACAATGTTGATGTTCATGATATAGAATTGTTACAGTATATCAATGTGGATTG 
               
               
                   
               
               
                 TGCAAAATTAAAACGACTCCTGAAGGAAACAGCATTTAAATTTAAAGCCCTAAAGAAGGTTGCGCAGTTA 
               
               
                   
               
               
                 GCAGTTATAAATAGCCTGGAAAAG   GCATTTTGGAACTGGGTAGAA AATTATCCAGATGAATTTACAAAAC   
               
               
                   
               
               
                   TGTACCAGATCCCACAGACTGATATGGCTG AATGTGCAGAAAAGCTATTTGACTTGGTGGATGGTTTTGC 
               
               
                   
               
               
                 TGAAAGCACCAAACGTAAAGCAGCAGTTTGGCCACTACAAATCATTCTCCTTATCTTGTGTCCAGAAATA 
               
               
                   
               
               
                 ATCCAGGATATATCCAAAGACGTGGTTGATGAAAACAACATGAATAAGAAGTTATTTCTGGACAGTCTAC 
               
               
                   
               
               
                 GAAAAGCTCTTGCTGGCCATGGAGGAAGTAGGCAGCTGACAGAAAGTGCTGCAATTGCCTGTGTCAAACT 
               
               
                   
               
               
                 GTGTAAAGCAAGTACTTACATCAATTGGGAAGATAACTCTGTCATTTTCCTAC   TTGTTCAGTCCATGGTG     
               
               
                   
               
               
                     GTT   GATCTTAAGAACCTGCTTTTTAATCCAAGTAAGCCATTCTCAAGAGGCAGTCAGCCTGCAGATGTGG 
               
               
                   
               
               
                 ATCTAATGATTGACTGCCTTGTTTCTTGCTTTCGTATAAGCCCTCACAACAACCAACACTTTAAGATCTG 
               
               
                   
               
               
                 CCTGGCTCAGAATTCACCTTCTACATTTCACTATGTGCTGGTAAATTCACTCCATCGAATCATCACCAAT 
               
               
                   
               
               
                 TCCGCATTGGATTGGTGGCCTAAGATTGATGCTGTGTATTGTCACTCGGTTGAACTTCGAAATATGTTTG 
               
               
                   
               
               
                 GTGAAACACTTCATAAAGCAGTGCAAGGTTGTGGAGCACACCCAGCAATACGAATGGCACCGAGTCTTAC 
               
               
                   
               
               
                 ATTTAAAGAAAAAGTAACAAGCCTTAAATTTAAAGAAAAACCTACAGACCTGGAGACAAGAAGCTATAAG 
               
               
                   
               
               
                 TATCTTCTCTTGTCCATGGTGAAACTAATTCATGCAGATCCAAAGCTCTTGCTTTGTAATCCAAGAAAAC 
               
               
                   
               
               
                 AGGGGCCCGAAACCCAAGGCAGTACAGCAGAATTAATTACAGGGCTCGTCCAACTGGTCCCTCAGTCACA 
               
               
                   
               
               
                 CATGCCAGAGATTGCTCAGGAAGCAATGGAGGCTCTGCTGGTTCTTCATCAGTTAGATAGCATTGATTTG 
               
               
                   
               
               
                 TGGAATCCTGATGCTCCTGTAGAAACATTTTGGGAGATTAGCTCACAAATGCTTTTTTACATCTGCAAGA 
               
               
                   
               
               
                 AATTAACTAGTCATCAAATGCTTAGTAGCACAGAAATTCTCAAGTGGTTGCGGGAAATATTGATCTGCAG 
               
               
                   
               
               
                 GAATAAATTTCTTCTTAAAAATAAGCAGGCAGATAGAAGTTCCTGTCACTTTCTCCTTTTTTACGGGGTA 
               
               
                   
               
               
                 GGATGTGATATTCCTTCTAGTGGAAATACCAGTCAAATGTCCATGGATCATGAAGAATTACTACGTACTC 
               
               
                   
               
               
                 CTGGAGCCTCTCTCCGGAAGGGAAAAGGGAACTCCTCTATGGATAGTGCAGCAGGATGCAGCGGAACCCC 
               
               
                   
               
               
                 CCCGATTTGCCGACAAGCCCAGACCAAACTAGAAGTGGCCCTGTACATGTTTCTGTGGAACCCTGACACT 
               
               
                   
               
               
                 GAAGCTGTTCTGGTTGCCATGTCCTGTTTCCGCCACCTCTGTGAGGAAGCAGATATCCGGTGTGGGGTGG 
               
               
                   
               
               
                 ATGAAGTGTCAGTGCATAACCTCTTGCCCAACTATAACACATTCATGGAGTTTGCCTCTGTCAGCAATAT 
               
               
                   
               
               
                 GATGTCAACAGGAAGAGCAGCACTTCAGAAAAGAGTGATGGCACTGCTGAGGCGCATTGAGCATCCCACT 
               
               
                   
               
               
                 GCAGGAAACACTGAGGCTTGGGAAGATACACATGCAAAATGGGAACAAGCAACAAAGCTAATCCTTAACT 
               
               
                   
               
               
                 ATCCAAAAGCCAAAATGGAAGATGGCCAGGCTGCTGAAAGCCTTCACAAGACCATTGTTAAGAGGCGAAT 
               
               
                   
               
               
                 GTCCCATGTGAGTGGAGGAGGATCCATAGATTTGTCTGACACAGACTCCCTACAGGAATGGATCAACATG 
               
               
                   
               
               
                 ACTGGCTTCCTTTGTGCCCTTGGGGGAGTGTGCCTCCAGCAGAGAAGCAATTCTGGCCTGGCAACCTATA 
               
               
                   
               
               
                 GCCCACCCATGGGTCCAGTCAGTGAACGTAAGGGTTCTATGATTTCAGTGATGTCTTCAGAGGGAAACGC 
               
               
                   
               
               
                 AGATACACCTGTCAGCAAATTTATGGATCGGCTGTTGTCCTTAATGGTGTGTAACCATGAGAAAGTGGGA 
               
               
                   
               
               
                 CTTCAAATACGGACCAATGTTAAGGATCTGGTGGGTCTAGAATTGAGTCCTGCTCTGTATCCAATGCTAT 
               
               
                   
               
               
                 TTAACAAATTGAAGAATACCATCAGCAAGTTTTTTGACTCCCAAGGACAGGTTTTATTGACTGATACCAA 
               
               
                   
               
               
                 TACTCAATTTGTAGAACAAACCATAGCTATAATGAAGAACTTGCTAGATAATCATACTGAAGGCAGCTCT 
               
               
                   
               
               
                 GAACATCTAGGGCAAGCTAGCATTGAAACAATGATGTTAAATCTGGTCAGGTATGTTCGTGTGCTTGGGA 
               
               
                   
               
               
                 ATATGGTCCATGCAATTCAAATAAAAACGAAACTGTGTCAATTAGTTGAAGTAATGATGGCAAGGAGAGA 
               
               
                   
               
               
                 TGACCTCTCATTTTGCCAAGAGATGAAATTTAGGAATAAGATGGTAGAATACCTGACAGACTGGGTTATG 
               
               
                   
               
               
                 GGAACATCAAACCAAGCAGCAGATGATGATGTAAAATGTCTTACAAGAGATTTGGACCAGGCAAGCATGG 
               
               
                   
               
               
                 AAGCAGTAGTTTCACTTCTAGCTGGTCTCCCTCTGCAGCCTGAAGAAGGAGATGGTGTGGAATTGATGGA 
               
               
                   
               
               
                 AGCCAAATCACAGTTATTTCTTAAATACTTCACATTATTTATGAACCTTTTGAATGACTGCAGTGAAGTT 
               
               
                   
               
               
                 GAAGATGAAAGTGCGCAAACAGGTGGCAGGAAACGTGGCATGTCTCGGAGGCTGGCATCACTGAGGCACT 
               
               
                   
               
               
                 GTACGGTCCTTGCAATGTCAAACTTACTCAATGCCAACGTAGACAGTGGTCTCATGCACTCCATAGGCTT 
               
               
                   
               
               
                 AGGTTACCACAAGGATCTCCAGACAAGAGCTACATTTATGGAAGTTCTGACAAAAATCCTTCAACAAGGC 
               
               
                   
               
               
                 ACAGAATTTGACACACTTGCAGAAACAGTATTGGCTGATCGGTTTGAGAGATTGGTGGAACTGGTCACAA 
               
               
                   
               
               
                 TGATGGGTGATCAAGGAGAACTCCCTATAGCGATGGCTCTGGCCAATGTGGTTCCTTGTTCTCAGTGGGA 
               
               
                   
               
               
                 TGAACTAGCTCGAGTTCTGGTTACTCTGTTTGATTCTCGGCATTTACTCTACCAACTGCTCTGGAACATG 
               
               
                   
               
               
                 TTTTCTAAAGAAGTAGAATTGGCAGACTCCATGCAGACTCTCTTCCGAGGCAACAGCTTGGCCAGTAAAA 
               
               
                   
               
               
                 TAATGACATTCTGTTTCAAGGTATATGGTGCTACCTATCTACAAAAACTCCTGGATCCTTTATTACGAAT 
               
               
                   
               
               
                 TGTGATCACATCCTCTGATTGGCAACATGTTAGCTTTGAAGTGGATCCTACCAGGTTAGAACCATCAGAG 
               
               
                   
               
               
                 AGCCTTGAGGAAAACCAGCGGAACCTCCTTCAGATGACTGAAAAGTTCTTCCATGCCATCATCAGTTCCT 
               
               
                   
               
               
                 CCTCAGAATTCCCCCCTCAACTTCGAAGTGTGTGCCACTGTTTATACCAGGCAACTTGCCACTCCCTACT 
               
               
                   
               
               
                 GAATAAAGCTACAGTAAAAGAAAAAAAGGAAAACAAAAAATCAGTGGTTAGCCAGCGTTTCCCTCAGAAC 
               
               
                   
               
               
                 AGCATCGGTGCAGTAGGAAGTGCCATGTTCCTCAGATTTATCAATCCTGCCATTGTCTCACCGTATGAAG 
               
               
                   
               
               
                 CAGGGATTTTAGATAAAAAGCCACCACCTAGAATCGAAAGGGGCTTGAAGTTAATGTCAAAGATACTTCA 
               
               
                   
               
               
                 GAGTATTGCCAATCATGTTCTCTTCACAAAAGAAGAACATATGCGGCCTTTCAATGATTTTGTGAAAAGC 
               
               
                   
               
               
                 AACTTTGATGCAGCACGCAGGTTTTTCCTTGATATAGCATCTGATTGTCCTACAAGTGATGCAGTAAATC 
               
               
                   
               
               
                 ATAGTCTTTCCTTCATAAGTGACGGCAATGTGCTTGCTTTACATCGTCTACTCTGGAACAATCAGGAGAA 
               
               
                   
               
               
                 AATTGGGCAGTATCTTTCCAGCAACAGGGATCATAAAGCTGTTGGAAGACGACCTTTTGATAAGATGGCA 
               
               
                   
               
               
                 ACACTTCTTGCATACCTGGGTCCTCCAGAGCACAAACCTGTGGCAGATACACACTGGTCCAGCCTTAACC 
               
               
                   
               
               
                 TTACCAGTTCAAAGTTTGAGGAATTTATGACTAGGCATCAGGTACATGAAAAAGAAGAATTCAAGGCTTT 
               
               
                   
               
               
                 GAAAACGTTAAGTATTTTCTACCAAGCTGGGACTTCCAAAGCTGGGAATCCTATTTTTTATTATGTTGCA 
               
               
                   
               
               
                 CGGAGGTTCAAAACTGGTCAAATCAATGGTGATTTGCTGATATACCATGTCTTACTGACTTTAAAGCCAT 
               
               
                   
               
               
                 ATTATGCAAAGCCATATGAAATTGTAGTGGACCTTACCCATACCGGGCCTAGCAATCGCTTTAAAACAGA 
               
               
                   
               
               
                 CTTTCTCTCTAAGTGGTTTGTTGTTTTTCCTGGCTTTGCTTACGACAACGTCTCCGCAGTCTATATCTAT 
               
               
                   
               
               
                 AACTGTAACTCCTGGGTCAGGGAGTACACCAAGTATCATGAGCGGCTGCTGACTGGCCTCAAAGGTAGCA 
               
               
                   
               
               
                 AAAGGCTTGTTTTCATAGACTGTCCTGGGAAACTGGCTGAGCACATAGAGCATGAACAACAGAAACTACC 
               
               
                   
               
               
                 TGCTGCCACCTTGGCTTTAGAAGAGGACCTGAAGGTATTCCACAATGCTCTCAAGCTAGCTCACAAAGAC 
               
               
                   
               
               
                 ACCAAAGTTTCTATTAAAGTTGGTTCTACTGCTGTCCAAGTAACTTCAGCAGAGCGAACAAAAGTCCTAG 
               
               
                   
               
               
                 GGCAATCAGTCTTTCTAAATGACATTTATTATGCTTCGGAAATTGAAGAAATCTGCCTAGTAGATGAGAA 
               
               
                   
               
               
                 CCAGTTCACCTTAACCATTGCAAACCAGGGCACGCCGCTCACCTTCATGCACCAGGAGTGTGAAGCCATT 
               
               
                   
               
               
                 GTCCAGTCTATCATTCATATCCGGACCCGCTGGGAACTGTCACAGCCCGACTCTATCCCCCAACACACCA 
               
               
                   
               
               
                 AGATTCGGCCAAAAGATGTCCCTGGGACACTGCTCAATATCGCATTACTTAATTTAGGCAGTTCTGACCC 
               
               
                   
               
               
                 GAGTTTACGGTCAGCTGCCTATAATCTTCTGTGTGCCTTAACTTGTACCTTTAATTTAAAAATCGAGGGC 
               
               
                   
               
               
                 CAGTTACTAGAGACATCAGGTTTATGTATCCCTGCCAACAACACCCTCTTTATTGTCTCTATTAGTAAGA 
               
               
                   
               
               
                 CACTGGCAGCCAATGAGCCACACCTCACGTTAGAATTTTTGGAAGAGTGTATTTCTGGATTTAGCAAATC 
               
               
                   
               
               
                 TAGTATTGAATTGAAACACCTTTGTTTGGAATACATGACTCCATGGCTGTCAAATCTAGTTCGTTTTTGC 
               
               
                   
               
               
                 AAGCATAATGATGATGCCAAACGACAAAGAGTTACTGCTATTCTTGACAAGCTGATAACAATGACCATCA 
               
               
                   
               
               
                 ATGAAAAACAGATGTACCCATCTATTCAAGCAAAAATATGGGGAAGCCTTGGGCAGATTACAGATCTGCT 
               
               
                   
               
               
                 TGATGTTGTACTAGACAGTTTCATCAAAACCAGTGCAACAGGTGGCTTGGGATCAATAAAAGCTGAGGTG 
               
               
                   
               
               
                 ATGGCAGATACTGCTGTAGCTTTGGCTTCTGGAAATGTGAAATTGGTTTCAAGCAAGGTTATTGGAAGGA 
               
               
                   
               
               
                 TGTGCAAAATAATTGACAAGACATGCTTATCTCCAACTCCTACTTTAGAACAACATCTTATGTGGGATGA 
               
               
                   
               
               
                 TATTGCTATTTTAGCACGCTACATGCTGATGCTGTCCTTCAACAATTCCCTTGATGTGGCAGCTCATCTT 
               
               
                   
               
               
                 CCCTACCTCTTCCACGTTGTTACTTTCTTAGTAGCCACAGGTCCGCTCTCCCTTAGAGCTTCCACACATG 
               
               
                   
               
               
                 GACTGGTCATTAATATCATTCACTCTCTGTGTACTTGTTCACAGCTTCATTTTAGTGAAGAGACCAAGCA 
               
               
                   
               
               
                 AGTTTTGAGACTCAGTCTGACAGAGTTCTCATTACCCAAATTTTACTTGCTGTTTGGCATTAGCAAAGTC 
               
               
                   
               
               
                 AAGTCAGCTGCTGTCATTGCCTTCCGTTCCAGTTACCGGGACAGGTCATTCTCTCCTGGCTCCTATGAGA 
               
               
                   
               
               
                 GAGAGACTTTTGCTTTGACATCCTTGGAAACAGTCACAGAAGCTTTGTTGGAGATCATGGAGGCATGCAT 
               
               
                   
               
               
                 GAGAGATATTCCAACGTGCAAGTGGCTGGACCAGTGGACAGAACTAGCTCAAAGATTTGCATTCCAATAT 
               
               
                   
               
               
                 AATCCATCCCTGCAACCAAGAGCTCTTGTTGTCTTTGGGTGTATTAGCAAACGAGTGTCTCATGGGCAGA 
               
               
                   
               
               
                 TAAAGCAGATAATCCGTATTCTTAGCAAGGCACTTGAGAGTTGCTTAAAAGGACCTGACACTTACAACAG 
               
               
                   
               
               
                 TCAAGTTCTGATAGAAGCTACAGTAATAGCACTAACCAAATTACAGCCACTTCTTAATAAGGACTCGCCT 
               
               
                   
               
               
                 CTGCACAAAGCCCTCTTTTGGGTAGCTGTGGCTGTGCTGCAGCTTGATGAGGTCAACTTGTATTCAGCAG 
               
               
                   
               
               
                 GTACCGCACTTCTTGAACAAAACCTGCATACTTTAGATAGTCTCCGTATATTCAATGACAAGAGTCCAGA 
               
               
                   
               
               
                 GGAAGTATTTATGGCAATCCGGAATCCTCTGGAGTGGCACTGCAAGCAAATGGATCATTTTGTTGGACTC 
               
               
                   
               
               
                 AATTTCAACTCTAACTTTAACTTTGCATTGGTTGGACACCTTTTAAAAGGGTACAGGCATCCTTCACCTG 
               
               
                   
               
               
                 CTATTGTTGCAAGAACAGTCAGAATTTTACATACACTACTAACTCTGGTTAACAAACACAGAAATTGTGA 
               
               
                   
               
               
                 CAAATTTGAAGTGAATACACAGAGCGTGGCCTACTTAGCAGCTTTACTTACAGTGTCTGAAGAAGTTCGA 
               
               
                   
               
               
                 AGTCGCTGCAGCCTAAAACATAGAAAGTCACTTCTTCTTACTGATATTTCAATGGAAAATGTTCCTATGG 
               
               
                   
               
               
                 ATACATATCCCATTCATCATGGTGACCCTTCCTATAGGACACTAAAGGAGACTCAGCCATGGTCCTCTCC 
               
               
                   
               
               
                 CAAAGGTTCTGAAGGATACCTTGCAGCCACCTATCCAACTGTCGGCCAGACCAGTCCCCGAGCCAGGAAA 
               
               
                   
               
               
                 TCCATGAGCCTGGACATGGGGCAACCTTCTCAGGCCAACACTAAGAAGTTGCTTGGAACAAGGAAAAGTT 
               
               
                   
               
               
                 TTGATCACTTGATATCAGACACAAAGGCTCCTAAAAGGCAAGAAATGGAATCAGGGATCACAACACCCCC 
               
               
                   
               
               
                 CAAAATGAGGAGAGTAGCAGAAACTGATTATGAAATGGAAACTCAGAGGATTTCCTCATCACAACAGCAC 
               
               
                   
               
               
                 CCACATTTACGTAAAGTTTCAGTGTCTGAATCAAATGTTCTCTTGGATGAAGAAGTACTTACTGATCCGA 
               
               
                   
               
               
                 AGATCCAGGCGCTGCTTCTTACTGTTCTAGCTACACTGGTAAAATATACCACAGATGAGTTTGATCAACG 
               
               
                   
               
               
                 AATTCTTTATGAATACTTAGCAGAGGCCAGTGTTGTGTTTCCCAAAGTCTTTCCTGTTGTGCATAATTTG 
               
               
                   
               
               
                 TTGGACTCTAAGATCAACACCCTGTTATCATTGTGCCAAGATCCAAATTTGTTAAATCCAATCCATGGAA 
               
               
                   
               
               
                 TTGTGCAGAGTGTGGTGTACCATGAAGAATCCCCACCACAATACCAAACATCTTACCTGCAAAGTTTTGG 
               
               
                   
               
               
                 TTTTAATGGCTTGTGGCGGTTTGCAGGACCGTTTTCAAAGCAAACACAAATTCCAGACTATGCTGAGCTT 
               
               
                   
               
               
                 ATTGTTAAGTTTCTTGATGCCTTGATTGACACGTACCTGCCTGGAATTGATGAAGAAACCAGTGAAGAAT 
               
               
                   
               
               
                 CCCTCCTGACTCCCACATCTCCTTACCCTCCTGCACTGCAGAGCCAGCTTAGTATCACTGCCAACCTTAA 
               
               
                   
               
               
                 CCTTTCTAATTCCATGACCTCACTTGCAACTTCCCAGCATTCCCCAGGAATCGACAAGGAGAACGTTGAA 
               
               
                   
               
               
                 CTCTCCCCTACCACTGGCCACTGTAACAGTGGACGAACTCGCCACGGATCCGCAAGCCAAGTGCAGAAGC 
               
               
                   
               
               
                 AAAGAAGCGCTGGCAGTTTCAAACGTAATAGCATTAAGAAGATCGTGTGA 
               
               
                   
               
               
                 (Exon 8 is indicated as double underline.) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 31 
               
               
                   
               
               
                 NF1 variant (lacking exon 8) Nucleotide Sequence 
               
               
                 (8444 nt, SEQ ID No. 76) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGGCCGCGCACAGGCCGGTGGAATGGGTCCAGGCCGTGGTCAGCCGCTTCGACGAGCAGCTTCCAATAA 
               
               
                   
               
               
                 AAACAGGACAGCAGAACACACATACCAAAGTCAGTACTGAGCACAACAAGGAATGTCTAATCAATATTTC 
               
               
                   
               
               
                 CAAATACAAGTTTTCTTTGGTTATAAGCGGCCTCACTACTATTTTAAAGAATGTTAACAATATGAGAATA 
               
               
                   
               
               
                 TTTGGAGAAGCTGCTGAAAAAAATTTATATCTCTCTCAGTTGATTATATTGGATACACTGGAAAAATGTC 
               
               
                   
               
               
                 TTGCTGGGCAACCAAAGGACACAATGAGATTAGATGAAACGATGCTGGTCAAACAGTTGCTGCCAGAAAT 
               
               
                   
               
               
                 CTGCCATTTTCTTCACACCTGTCGTGAAGGAAACCAGCATGCAGCTGAACTTCGGAATTCTGCCTCTGGG 
               
               
                   
               
               
                 GTTTTATTTTCTCTCAGCTGCAACAACTTCAATGCAGTCTTTAGTCGCATTTCTACCAGGTTACAGGAAT 
               
               
                   
               
               
                 TAACTGTTTGTTCAGAAGACAATGTTGATGTTCATGATATAGAATTGTTACAGTATATCAATGTGGATTG 
               
               
                   
               
               
                 TGCAAAATTAAAACGACTCCTGAAGGAAACAGCATTTAAATTTAAAGCCCTAAAGAAGGTTGCGCAGTTA 
               
               
                   
               
               
                 GCAGTTATAAATA   GCCTGGAAAA       GA       ATGTGCAGA   AAAGCTATTTGACTTGGTGGATGGTTTTGCTGAAAG 
               
               
                   
               
               
                 CACCAAACGTAAAGCAGCAGTTTGGCCACTACAAATCATTCTCCTTATCTTGTGTCCAGAAATAATCCAG 
               
               
                   
               
               
                 GATATATCCAAAGACGTGGTTGATGAAAACAACATGAATAAGAAGTTATTTCTGGACAGTCTACGAAAAG 
               
               
                   
               
               
                 CTCTTGCTGGCCATGGAGGAAGTAGGCAGCTGACAGAAAGTGCTGCAATTGCCTGTGTCAAACTGTGTAA 
               
               
                   
               
               
                 AGCAAGTACTTACATCAATTGGGAAGATAACTCTGTCATTTTCCTACTTGTTCAGTCCATGGTGGTTGAT 
               
               
                   
               
               
                 CTTAAGAACCTGCTTTTTAATCCAAGTAAGCCATTCTCAAGAGGCAGTCAGCCTGCAGATGTGGATCTAA 
               
               
                   
               
               
                 TGATTGACTGCCTTGTTTCTTGCTTTCGTATAAGCCCTCACAACAACCAACACTTTAAGATCTGCCTGGC 
               
               
                   
               
               
                 TCAGAATTCACCTTCTACATTTCACTATGTGCTGGTAAATTCACTCCATCGAATCATCACCAATTCCGCA 
               
               
                   
               
               
                 TTGGATTGGTGGCCTAAGATTGATGCTGTGTATTGTCACTCGGTTGAACTTCGAAATATGTTTGGTGAAA 
               
               
                   
               
               
                 CACTTCATAAAGCAGTGCAAGGTTGTGGAGCACACCCAGCAATACGAATGGCACCGAGTCTTACATTTAA 
               
               
                   
               
               
                 AGAAAAAGTAACAAGCCTTAAATTTAAAGAAAAACCTACAGACCTGGAGACAAGAAGCTATAAGTATCTT 
               
               
                   
               
               
                 CTCTTGTCCATGGTGAAACTAATTCATGCAGATCCAAAGCTCTTGCTTTGTAATCCAAGAAAACAGGGGC 
               
               
                   
               
               
                 CCGAAACCCAAGGCAGTACAGCAGAATTAATTACAGGGCTCGTCCAACTGGTCCCTCAGTCACACATGCC 
               
               
                   
               
               
                 AGAGATTGCTCAGGAAGCAATGGAGGCTCTGCTGGTTCTTCATCAGTTAGATAGCATTGATTTGTGGAAT 
               
               
                   
               
               
                 CCTGATGCTCCTGTAGAAACATTTTGGGAGATTAGCTCACAAATGCTTTTTTACATCTGCAAGAAATTAA 
               
               
                   
               
               
                 CTAGTCATCAAATGCTTAGTAGCACAGAAATTCTCAAGTGGTTGCGGGAAATATTGATCTGCAGGAATAA 
               
               
                   
               
               
                 ATTTCTTCTTAAAAATAAGCAGGCAGATAGAAGTTCCTGTCACTTTCTCCTTTTTTACGGGGTAGGATGT 
               
               
                   
               
               
                 GATATTCCTTCTAGTGGAAATACCAGTCAAATGTCCATGGATCATGAAGAATTACTACGTACTCCTGGAG 
               
               
                   
               
               
                 CCTCTCTCCGGAAGGGAAAAGGGAACTCCTCTATGGATAGTGCAGCAGGATGCAGCGGAACCCCCCCGAT 
               
               
                   
               
               
                 TTGCCGACAAGCCCAGACCAAACTAGAAGTGGCCCTGTACATGTTTCTGTGGAACCCTGACACTGAAGCT 
               
               
                   
               
               
                 GTTCTGGTTGCCATGTCCTGTTTCCGCCACCTCTGTGAGGAAGCAGATATCCGGTGTGGGGTGGATGAAG 
               
               
                   
               
               
                 TGTCAGTGCATAACCTCTTGCCCAACTATAACACATTCATGGAGTTTGCCTCTGTCAGCAATATGATGTC 
               
               
                   
               
               
                 AACAGGAAGAGCAGCACTTCAGAAAAGAGTGATGGCACTGCTGAGGCGCATTGAGCATCCCACTGCAGGA 
               
               
                   
               
               
                 AACACTGAGGCTTGGGAAGATACACATGCAAAATGGGAACAAGCAACAAAGCTAATCCTTAACTATCCAA 
               
               
                   
               
               
                 AAGCCAAAATGGAAGATGGCCAGGCTGCTGAAAGCCTTCACAAGACCATTGTTAAGAGGCGAATGTCCCA 
               
               
                   
               
               
                 TGTGAGTGGAGGAGGATCCATAGATTTGTCTGACACAGACTCCCTACAGGAATGGATCAACATGACTGGC 
               
               
                   
               
               
                 TTCCTTTGTGCCCTTGGGGGAGTGTGCCTCCAGCAGAGAAGCAATTCTGGCCTGGCAACCTATAGCCCAC 
               
               
                   
               
               
                 CCATGGGTCCAGTCAGTGAACGTAAGGGTTCTATGATTTCAGTGATGTCTTCAGAGGGAAACGCAGATAC 
               
               
                   
               
               
                 ACCTGTCAGCAAATTTATGGATCGGCTGTTGTCCTTAATGGTGTGTAACCATGAGAAAGTGGGACTTCAA 
               
               
                   
               
               
                 ATACGGACCAATGTTAAGGATCTGGTGGGTCTAGAATTGAGTCCTGCTCTGTATCCAATGCTATTTAACA 
               
               
                   
               
               
                 AATTGAAGAATACCATCAGCAAGTTTTTTGACTCCCAAGGACAGGTTTTATTGACTGATACCAATACTCA 
               
               
                   
               
               
                 ATTTGTAGAACAAACCATAGCTATAATGAAGAACTTGCTAGATAATCATACTGAAGGCAGCTCTGAACAT 
               
               
                   
               
               
                 CTAGGGCAAGCTAGCATTGAAACAATGATGTTAAATCTGGTCAGGTATGTTCGTGTGCTTGGGAATATGG 
               
               
                   
               
               
                 TCCATGCAATTCAAATAAAAACGAAACTGTGTCAATTAGTTGAAGTAATGATGGCAAGGAGAGATGACCT 
               
               
                   
               
               
                 CTCATTTTGCCAAGAGATGAAATTTAGGAATAAGATGGTAGAATACCTGACAGACTGGGTTATGGGAACA 
               
               
                   
               
               
                 TCAAACCAAGCAGCAGATGATGATGTAAAATGTCTTACAAGAGATTTGGACCAGGCAAGCATGGAAGCAG 
               
               
                   
               
               
                 TAGTTTCACTTCTAGCTGGTCTCCCTCTGCAGCCTGAAGAAGGAGATGGTGTGGAATTGATGGAAGCCAA 
               
               
                   
               
               
                 ATCACAGTTATTTCTTAAATACTTCACATTATTTATGAACCTTTTGAATGACTGCAGTGAAGTTGAAGAT 
               
               
                   
               
               
                 GAAAGTGCGCAAACAGGTGGCAGGAAACGTGGCATGTCTCGGAGGCTGGCATCACTGAGGCACTGTACGG 
               
               
                   
               
               
                 TCCTTGCAATGTCAAACTTACTCAATGCCAACGTAGACAGTGGTCTCATGCACTCCATAGGCTTAGGTTA 
               
               
                   
               
               
                 CCACAAGGATCTCCAGACAAGAGCTACATTTATGGAAGTTCTGACAAAAATCCTTCAACAAGGCACAGAA 
               
               
                   
               
               
                 TTTGACACACTTGCAGAAACAGTATTGGCTGATCGGTTTGAGAGATTGGTGGAACTGGTCACAATGATGG 
               
               
                   
               
               
                 GTGATCAAGGAGAACTCCCTATAGCGATGGCTCTGGCCAATGTGGTTCCTTGTTCTCAGTGGGATGAACT 
               
               
                   
               
               
                 AGCTCGAGTTCTGGTTACTCTGTTTGATTCTCGGCATTTACTCTACCAACTGCTCTGGAACATGTTTTCT 
               
               
                   
               
               
                 AAAGAAGTAGAATTGGCAGACTCCATGCAGACTCTCTTCCGAGGCAACAGCTTGGCCAGTAAAATAATGA 
               
               
                   
               
               
                 CATTCTGTTTCAAGGTATATGGTGCTACCTATCTACAAAAACTCCTGGATCCTTTATTACGAATTGTGAT 
               
               
                   
               
               
                 CACATCCTCTGATTGGCAACATGTTAGCTTTGAAGTGGATCCTACCAGGTTAGAACCATCAGAGAGCCTT 
               
               
                   
               
               
                 GAGGAAAACCAGCGGAACCTCCTTCAGATGACTGAAAAGTTCTTCCATGCCATCATCAGTTCCTCCTCAG 
               
               
                   
               
               
                 AATTCCCCCCTCAACTTCGAAGTGTGTGCCACTGTTTATACCAGGCAACTTGCCACTCCCTACTGAATAA 
               
               
                   
               
               
                 AGCTACAGTAAAAGAAAAAAAGGAAAACAAAAAATCAGTGGTTAGCCAGCGTTTCCCTCAGAACAGCATC 
               
               
                   
               
               
                 GGTGCAGTAGGAAGTGCCATGTTCCTCAGATTTATCAATCCTGCCATTGTCTCACCGTATGAAGCAGGGA 
               
               
                   
               
               
                 TTTTAGATAAAAAGCCACCACCTAGAATCGAAAGGGGCTTGAAGTTAATGTCAAAGATACTTCAGAGTAT 
               
               
                   
               
               
                 TGCCAATCATGTTCTCTTCACAAAAGAAGAACATATGCGGCCTTTCAATGATTTTGTGAAAAGCAACTTT 
               
               
                   
               
               
                 GATGCAGCACGCAGGTTTTTCCTTGATATAGCATCTGATTGTCCTACAAGTGATGCAGTAAATCATAGTC 
               
               
                   
               
               
                 TTTCCTTCATAAGTGACGGCAATGTGCTTGCTTTACATCGTCTACTCTGGAACAATCAGGAGAAAATTGG 
               
               
                   
               
               
                 GCAGTATCTTTCCAGCAACAGGGATCATAAAGCTGTTGGAAGACGACCTTTTGATAAGATGGCAACACTT 
               
               
                   
               
               
                 CTTGCATACCTGGGTCCTCCAGAGCACAAACCTGTGGCAGATACACACTGGTCCAGCCTTAACCTTACCA 
               
               
                   
               
               
                 GTTCAAAGTTTGAGGAATTTATGACTAGGCATCAGGTACATGAAAAAGAAGAATTCAAGGCTTTGAAAAC 
               
               
                   
               
               
                 GTTAAGTATTTTCTACCAAGCTGGGACTTCCAAAGCTGGGAATCCTATTTTTTATTATGTTGCACGGAGG 
               
               
                   
               
               
                 TTCAAAACTGGTCAAATCAATGGTGATTTGCTGATATACCATGTCTTACTGACTTTAAAGCCATATTATG 
               
               
                   
               
               
                 CAAAGCCATATGAAATTGTAGTGGACCTTACCCATACCGGGCCTAGCAATCGCTTTAAAACAGACTTTCT 
               
               
                   
               
               
                 CTCTAAGTGGTTTGTTGTTTTTCCTGGCTTTGCTTACGACAACGTCTCCGCAGTCTATATCTATAACTGT 
               
               
                   
               
               
                 AACTCCTGGGTCAGGGAGTACACCAAGTATCATGAGCGGCTGCTGACTGGCCTCAAAGGTAGCAAAAGGC 
               
               
                   
               
               
                 TTGTTTTCATAGACTGTCCTGGGAAACTGGCTGAGCACATAGAGCATGAACAACAGAAACTACCTGCTGC 
               
               
                   
               
               
                 CACCTTGGCTTTAGAAGAGGACCTGAAGGTATTCCACAATGCTCTCAAGCTAGCTCACAAAGACACCAAA 
               
               
                   
               
               
                 GTTTCTATTAAAGTTGGTTCTACTGCTGTCCAAGTAACTTCAGCAGAGCGAACAAAAGTCCTAGGGCAAT 
               
               
                   
               
               
                 CAGTCTTTCTAAATGACATTTATTATGCTTCGGAAATTGAAGAAATCTGCCTAGTAGATGAGAACCAGTT 
               
               
                   
               
               
                 CACCTTAACCATTGCAAACCAGGGCACGCCGCTCACCTTCATGCACCAGGAGTGTGAAGCCATTGTCCAG 
               
               
                   
               
               
                 TCTATCATTCATATCCGGACCCGCTGGGAACTGTCACAGCCCGACTCTATCCCCCAACACACCAAGATTC 
               
               
                   
               
               
                 GGCCAAAAGATGTCCCTGGGACACTGCTCAATATCGCATTACTTAATTTAGGCAGTTCTGACCCGAGTTT 
               
               
                   
               
               
                 ACGGTCAGCTGCCTATAATCTTCTGTGTGCCTTAACTTGTACCTTTAATTTAAAAATCGAGGGCCAGTTA 
               
               
                   
               
               
                 CTAGAGACATCAGGTTTATGTATCCCTGCCAACAACACCCTCTTTATTGTCTCTATTAGTAAGACACTGG 
               
               
                   
               
               
                 CAGCCAATGAGCCACACCTCACGTTAGAATTTTTGGAAGAGTGTATTTCTGGATTTAGCAAATCTAGTAT 
               
               
                   
               
               
                 TGAATTGAAACACCTTTGTTTGGAATACATGACTCCATGGCTGTCAAATCTAGTTCGTTTTTGCAAGCAT 
               
               
                   
               
               
                 AATGATGATGCCAAACGACAAAGAGTTACTGCTATTCTTGACAAGCTGATAACAATGACCATCAATGAAA 
               
               
                   
               
               
                 AACAGATGTACCCATCTATTCAAGCAAAAATATGGGGAAGCCTTGGGCAGATTACAGATCTGCTTGATGT 
               
               
                   
               
               
                 TGTACTAGACAGTTTCATCAAAACCAGTGCAACAGGTGGCTTGGGATCAATAAAAGCTGAGGTGATGGCA 
               
               
                   
               
               
                 GATACTGCTGTAGCTTTGGCTTCTGGAAATGTGAAATTGGTTTCAAGCAAGGTTATTGGAAGGATGTGCA 
               
               
                   
               
               
                 AAATAATTGACAAGACATGCTTATCTCCAACTCCTACTTTAGAACAACATCTTATGTGGGATGATATTGC 
               
               
                   
               
               
                 TATTTTAGCACGCTACATGCTGATGCTGTCCTTCAACAATTCCCTTGATGTGGCAGCTCATCTTCCCTAC 
               
               
                   
               
               
                 CTCTTCCACGTTGTTACTTTCTTAGTAGCCACAGGTCCGCTCTCCCTTAGAGCTTCCACACATGGACTGG 
               
               
                   
               
               
                 TCATTAATATCATTCACTCTCTGTGTACTTGTTCACAGCTTCATTTTAGTGAAGAGACCAAGCAAGTTTT 
               
               
                   
               
               
                 GAGACTCAGTCTGACAGAGTTCTCATTACCCAAATTTTACTTGCTGTTTGGCATTAGCAAAGTCAAGTCA 
               
               
                   
               
               
                 GCTGCTGTCATTGCCTTCCGTTCCAGTTACCGGGACAGGTCATTCTCTCCTGGCTCCTATGAGAGAGAGA 
               
               
                   
               
               
                 CTTTTGCTTTGACATCCTTGGAAACAGTCACAGAAGCTTTGTTGGAGATCATGGAGGCATGCATGAGAGA 
               
               
                   
               
               
                 TATTCCAACGTGCAAGTGGCTGGACCAGTGGACAGAACTAGCTCAAAGATTTGCATTCCAATATAATCCA 
               
               
                   
               
               
                 TCCCTGCAACCAAGAGCTCTTGTTGTCTTTGGGTGTATTAGCAAACGAGTGTCTCATGGGCAGATAAAGC 
               
               
                   
               
               
                 AGATAATCCGTATTCTTAGCAAGGCACTTGAGAGTTGCTTAAAAGGACCTGACACTTACAACAGTCAAGT 
               
               
                   
               
               
                 TCTGATAGAAGCTACAGTAATAGCACTAACCAAATTACAGCCACTTCTTAATAAGGACTCGCCTCTGCAC 
               
               
                   
               
               
                 AAAGCCCTCTTTTGGGTAGCTGTGGCTGTGCTGCAGCTTGATGAGGTCAACTTGTATTCAGCAGGTACCG 
               
               
                   
               
               
                 CACTTCTTGAACAAAACCTGCATACTTTAGATAGTCTCCGTATATTCAATGACAAGAGTCCAGAGGAAGT 
               
               
                   
               
               
                 ATTTATGGCAATCCGGAATCCTCTGGAGTGGCACTGCAAGCAAATGGATCATTTTGTTGGACTCAATTTC 
               
               
                   
               
               
                 AACTCTAACTTTAACTTTGCATTGGTTGGACACCTTTTAAAAGGGTACAGGCATCCTTCACCTGCTATTG 
               
               
                   
               
               
                 TTGCAAGAACAGTCAGAATTTTACATACACTACTAACTCTGGTTAACAAACACAGAAATTGTGACAAATT 
               
               
                   
               
               
                 TGAAGTGAATACACAGAGCGTGGCCTACTTAGCAGCTTTACTTACAGTGTCTGAAGAAGTTCGAAGTCGC 
               
               
                   
               
               
                 TGCAGCCTAAAACATAGAAAGTCACTTCTTCTTACTGATATTTCAATGGAAAATGTTCCTATGGATACAT 
               
               
                   
               
               
                 ATCCCATTCATCATGGTGACCCTTCCTATAGGACACTAAAGGAGACTCAGCCATGGTCCTCTCCCAAAGG 
               
               
                   
               
               
                 TTCTGAAGGATACCTTGCAGCCACCTATCCAACTGTCGGCCAGACCAGTCCCCGAGCCAGGAAATCCATG 
               
               
                   
               
               
                 AGCCTGGACATGGGGCAACCTTCTCAGGCCAACACTAAGAAGTTGCTTGGAACAAGGAAAAGTTTTGATC 
               
               
                   
               
               
                 ACTTGATATCAGACACAAAGGCTCCTAAAAGGCAAGAAATGGAATCAGGGATCACAACACCCCCCAAAAT 
               
               
                   
               
               
                 GAGGAGAGTAGCAGAAACTGATTATGAAATGGAAACTCAGAGGATTTCCTCATCACAACAGCACCCACAT 
               
               
                   
               
               
                 TTACGTAAAGTTTCAGTGTCTGAATCAAATGTTCTCTTGGATGAAGAAGTACTTACTGATCCGAAGATCC 
               
               
                   
               
               
                 AGGCGCTGCTTCTTACTGTTCTAGCTACACTGGTAAAATATACCACAGATGAGTTTGATCAACGAATTCT 
               
               
                   
               
               
                 TTATGAATACTTAGCAGAGGCCAGTGTTGTGTTTCCCAAAGTCTTTCCTGTTGTGCATAATTTGTTGGAC 
               
               
                   
               
               
                 TCTAAGATCAACACCCTGTTATCATTGTGCCAAGATCCAAATTTGTTAAATCCAATCCATGGAATTGTGC 
               
               
                   
               
               
                 AGAGTGTGGTGTACCATGAAGAATCCCCACCACAATACCAAACATCTTACCTGCAAAGTTTTGGTTTTAA 
               
               
                   
               
               
                 TGGCTTGTGGCGGTTTGCAGGACCGTTTTCAAAGCAAACACAAATTCCAGACTATGCTGAGCTTATTGTT 
               
               
                   
               
               
                 AAGTTTCTTGATGCCTTGATTGACACGTACCTGCCTGGAATTGATGAAGAAACCAGTGAAGAATCCCTCC 
               
               
                   
               
               
                 TGACTCCCACATCTCCTTACCCTCCTGCACTGCAGAGCCAGCTTAGTATCACTGCCAACCTTAACCTTTC 
               
               
                   
               
               
                 TAATTCCATGACCTCACTTGCAACTTCCCAGCATTCCCCAGGAATCGACAAGGAGAACGTTGAACTCTCC 
               
               
                   
               
               
                 CCTACCACTGGCCACTGTAACAGTGGACGAACTCGCCACGGATCCGCAAGCCAAGTGCAGAAGCAAAGAA 
               
               
                   
               
               
                 GCGCTGGCAGTTTCAAACGTAATAGCATTAAGAAGATCGTGTGA 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
               
             
           
               
                 TABLE 32 
               
               
                   
               
               
                 Primer across the junction between NF1 
               
               
                 exon 7 and 9 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Primer across the junction 
                 
                   
                     GCCTGGAAAA 
                   
                   
                     GA 
                   
                   
                     ATGTGCAGA 
                   
                 
               
               
                 between NF1 exon 7 and 9 
                   
               
               
                 (SEQ ID No. 75) 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
           
               
                 TABLE 33 
               
               
                   
               
               
                 siRNA for selectively knockdown NF1 
               
               
                 full length and variants expression 
               
               
                   
               
             
             
               
                 siRNA targeting NF1 exon 8 
               
               
                 Sense (SEQ ID No. 73) 5′ CCAGAUCCCACAGACUGAUdTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 74) 3′ dTdTGGUCUAGGGUGUCUGA- 
               
               
                 CUA (5′-P)5′ 
               
               
                   
               
               
                 siRNA targeting splice junction between 
               
               
                 NF1 exon 7 and exon 9 
               
               
                 Sense (SEQ ID No. 77) 5′ GGAAAA GA AUGUGCAGAAAdTdT 
               
               
                 3′ 
               
               
                   
               
               
                 Antisense (SEQ ID No. 78) 3′ dTdTCCUUUU CU UACACGUC- 
               
               
                 UUU(5′-P)5′ 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 34 
               
               
                   
               
               
                 BAK1 (full length)Nucleotide Sequence 
               
               
                 (636 nt, SEQ ID No. 79) 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGGCTTCGGGGCAAGGCCCAGGTCCTCCCAGGCAGGAGTGCGGAGAGCC 
               
               
                   
               
               
                 TGCCCTGCCCTCTGCTTCTG AGGAGCAGGTAGCCCAGGACACAGAGGAGG   
               
               
                   
               
               
                 
                   TTTTCCGCAGCTACGTTTTTTACCGCCATCAGCAGGAACAGGAGGCTGAA 
                 
               
               
                   
               
               
                 
                   GGGGTGGCTGCCCCTGCCGACCCAGAG ATGGTCACCTTACCTCTGCAA CC 
                 
               
               
                   
               
               
                   TAGCA GCACCATGGGGCAGGTGGGACGGCAGCTCGCCATCATCGGGGACG 
               
               
                   
               
               
                 ACATCAACCGACGCTATGACTCAGAGTTCCAGACCATGTTGCAGCACCTG 
               
               
                   
               
               
                 CAGCCCACGGCAGAGAATGCCTATGAGTACTTCACCAAGATTGCCACCAG 
               
               
                   
               
               
                     CCTGTTTGAGAGTGGCATCAA   TTGGGGCCGTGTGGTGGCTCTTCTGGGCT 
               
               
                   
               
               
                 TCGGCTACCGTCTGGCCCTACACGTCTACCAGCATGGCCTGACTGGCTTC 
               
               
                   
               
               
                 CTAGGCCAGGTGACCCGCTTCGTGGTCGACTTCATGCTGCATCACTGCAT 
               
               
                   
               
               
                 TGCCCGGTGGATTGCACAGAGGGGTGGCTGGGTGGCAGCCCTGAACTTGG 
               
               
                   
               
               
                 GCAATGGTCCCATCCTGAACGTGCTGGTGGTTCTGGGTGTGGTTCTGTTG 
               
               
                   
               
               
                 GGCCAGTTTGTGGTACGAAGATTCTTCAAATCATGA 
               
               
                   
               
               
                 (Exon 2 is indicated as double underline.) 
               
             
          
         
       
     
     
       
         
               
             
               
             
           
               
                 TABLE 35 
               
               
                   
               
               
                 BAK1 variant (lacking exon 2) Nucleotide 
               
               
                 Sequence (501 nt, SEQ ID No. 85). 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ATGGCTTCGGGGCAAGGCCCAGGTCCTCCCAGGCAGGAGTGCGGAGAGCC 
               
               
                   
               
               
                 TGCCCTGCCC TCTGCTTCT   GG   CACCATGGG GCAGGTGGGACGGCAGCTCG 
               
               
                   
               
               
                 CCATCATCGGGGACGACATCAACCGACGCTATGACTCAGAGTTCCAGACC 
               
               
                   
               
               
                 ATGTTGCAGCACCTGCAGCCCACGGCAGAGAATGCCTATGAGTACTTCAC 
               
               
                   
               
               
                 CAAGATTGCCACCAGCCTGTTTGAGAGTGGCATCAATTGGGGCCGTGTGG 
               
               
                   
               
               
                 TGGCTCTTCTGGGCTTCGGCTACCGTCTGGCCCTACACGTCTACCAGCAT 
               
               
                   
               
               
                 GGCCTGACTGGCTTCCTAGGCCAGGTGACCCGCTTCGTGGTCGACTTCAT 
               
               
                   
               
               
                 GCTGCATCACTGCATTGCCCGGTGGATTGCACAGAGGGGTGGCTGGGTGG 
               
               
                   
               
               
                 CAGCCCTGAACTTGGGCAATGGTCCCATCCTGAACGTGCTGGTGGTTCTG 
               
               
                   
               
               
                 GGTGTGGTTCTGTTGGGCCAGTTTGTGGTACGAAGATTCTTCAAATCATG 
               
               
                   
               
               
                 A 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
     
       
         
               
             
               
               
             
           
               
                 TABLE 36 
               
               
                   
               
               
                 Primer across the junction between BAK1 
               
               
                 exon 7 and 9 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Primer across the junction 
                 
                   
                     TCTGCTTCT 
                   
                   
                     GG 
                   
                   
                     CACCATGGG 
                   
                 
               
               
                 between BAK1 exon 1 and 3 
                   
               
               
                 (SEQ ID No. 84) 
               
               
                   
               
               
                 (Double underline indicates bases bordering the splice junction) 
               
             
          
         
       
     
                     TABLE 37               siRNA for selectively knockdown BAK1       full length and variants expression                   siRNA targeting exon 2       Sense (SEQ ID No. 82) 5′ GGUCACCUUACCUCUGCAAdTdT       3′               Antisense (SEQ ID No. 83) 3′ dTdTCCAGUGGAAUGGAGAC-       GUU(5′-P)5′               siRNA targeting splice junction between       exon 1 and exon 3       Sense (SEQ ID No. 86) 5′ CCCUCUGCUUCUGGCACCAdTdT       3′               Antisense (SEQ ID No. 87) 3′ dTdTGGGAGACGAAGACCGU-       GGU (5′-P)5′               (Double underline indicates bases bordering the splice junction)            
Methods of Detection
 
     The present invention provides a method of identifying splicing variants of genes associated with prostate cancer risk and survival. The method generally comprises detecting the splicing variants in a nucleic acid sample from an individual, such as a prostate biopsy specimen. Typically, total RNA is extracted from the specimen, cDNA is synthesized from the extracted RNA and subject to further analysis. Nucleic acid samples used in the methods and assays of the present invention may be prepared by any available method or process. 
     Detection of splicing variants may be accomplished by amplifying specific fragments directly from a cDNA preparation from the tumor tissue using PCR. Presence of certain PCR product can be indicative of the presence of certain splicing variants, when the primers for the PCR are designed in such way that PCR products are only available when certain variants are present in the sample. Alternatively, primers may be designed to produce easily differentiable products for different variants. The sequence composition of the variants may also be determined from the amplified product. 
     The PCR reaction is well known in the art (See, e.g., U.S. Pat. No. 4,683,203; and U.S. Pat. No. 4,683,195). In general, the PCR procedure describes a method of gene amplification which is comprised of (i) sequence-specific hybridization of primers to specific genes within a DNA sample (or library), (ii) subsequent amplification involving multiple rounds of annealing, elongation, and denaturation using a DNA polymerase, and (iii) screening the PCR products for a band of the correct size. The primers used are oligonucleotides of sufficient length and appropriate sequence to provide initiation of polymerization, i.e. each primer is specifically designed to be complementary to each strand of the genomic locus to be amplified. The primers are prepared using any suitable method, such as conventional phosphotriester or phosphodiester methods or automated embodiments thereof (Beaucage,  Tet. Lett.  22:1859-1862, 1981). 
     For the detection of splicing variants, primers may be designed to flank a certain exon that may be alternatively spliced, i.e., one primer is complementary to the 5′ side of the exon, and the other primer is complementary to the 3′ side of the exon. The PCR amplification products thus would show different sizes. When the exon is present, a larger amplification product is obtained. When the exon is absent, a smaller amplification product is obtained. Alternatively, a primer may be designed to be complementary to a nucleotide sequence within the exon. This way, PCR amplification product is only available when the exon is present in the specimen. Additionally, a primer may be designed to be partially complementary to the 3′ end of an exon 5′ to the alternatively spliced exon, and partially complementary to the 5′ end of an exon 3′ to the alternatively spliced exon. PCR amplification product can only be obtained when the alternatively spliced exon is present in the sample. 
     The polymerization agent can be any compound or system (including enzymes) which will facilitate combination of the nucleotides in the proper manner to form the primer extension products which are complementary to each nucleic acid strand. Other fundamental conditions to allow amplification include the presence of nucleoside triphosphates and suitable temperature and pH (Thigpen et al.,  J. Clin. Invest.  90: 799-809, 1992; Saiki et al.,  Science  239: 487-491, 1988). 
     DNA sequences of the specified gene which have been amplified by use of polymerase chain reaction may also be screened using exon oligonucleotide probes. These probes are nucleic acid oligomers, each of which are complementary to a corresponding segment of the investigated gene and may or may not contain a known variant. The assay is performed by detecting the presence or absence of a hybridization signal for the specific sequence. 
     Oligonucleotide Probes 
     Another aspect of the subject invention is to provide for variant specific nucleic acid hybridization probes capable of detecting splicing variants of genes which predispose an individual to prostate cancer. The hybridization probes of the subject invention may be derived from the disclosed nucleotide sequences of the identified variants and form stable hybrids with the target sequences, under stringent to moderately stringent hybridization and wash conditions. Stringent conditions will be used in the case of perfect complementation with the target sequence, less stringent hybridization conditions will be used if mismatches are expected among the variants. Conditions will always be chosen such that nonspecific/adventitious bindings are eliminated or minimized. The probes may be of any suitable length, which span all or a portion of the specified gene region, and which allow specific hybridization. 
     Nucleic acid hybridization simply involves contacting a probe and target nucleic acid (from a nucleic acid sample) under conditions where the probe and its complementary target can form stable hybrid duplexes through complementary base pairing (see U.S. Pat. No. 6,333,155). Methods of nucleic acid hybridization are well known in the art. In a preferred embodiment, the probes are immobilized on solid supports such as beads, microarrays, or gene chips. 
     The probes include an isolated polynucleotide, preferably attached to a label or reporter molecule, may be used to isolate other polynucleotide sequences, having sequence similarity by standard methods. Techniques for preparing and labeling probes are known in the art and disclosed in Sambrook et al. (Molecular Cloning: A Laboratory Manual, Ed. 2; Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory, 1989) or Ausubel et al. (Current Protocols in Molecular Biology, Wiley &amp; Sons, New York, N.Y., 1995). The labels may be incorporated by any of a number of means well known to those of skill in the art (see U.S. Pat. No. 6,333,155). Commonly employed labels include, but are not limited to, biotin, fluorescent molecules, radioactive molecules, chromogenic substrates, chemiluminescent labels, enzymes, and the like. The methods for biotinylating nucleic acids are well known in the art, as are methods for introducing fluorescent molecules and radioactive molecules into oligonucleotides and nucleotides. 
     Other similar polynucleotides may be selected by using homologous polynucleotides. Alternatively, polynucleotides encoding these or similar polypeptides may be synthesized or selected by use of the redundancy in the genetic code. Various codon substitutions may be introduced, e.g., by silent changes (thereby producing various restriction sites) or to optimize expression for a particular system. Mutations may be introduced to modify the properties of the polypeptide, perhaps to change ligand-binding affinities, interchain affinities, or the polypeptide degradation or turnover rate. 
     Probes comprising synthetic oligonucleotides or other polynucleotides of the present invention may be derived from naturally occurring or recombinant single- or double-stranded polynucleotides, or be chemically synthesized. Probes may also be labeled by nick translation, Klenow fill-in reaction, or other methods known in the art. 
     Other means for producing specific hybridization probes for nucleic acids include the cloning of nucleic acid sequences into vectors for the production of mRNA probes. Such vectors are known in the art and are commercially available and may be used to synthesize RNA probes in vitro by means of the addition of the appropriate RNA polymerase as T7 or SP6 RNA polymerase and the appropriate radioactively labeled nucleotides. 
     The nucleotide sequences may be used to construct hybridization probes for mapping their respective genomic sequences. The nucleotide sequence provided herein may be mapped to a chromosome or specific regions of a chromosome using well known genetic and/or chromosomal mapping techniques. These techniques include in situ hybridization, linkage analysis against known chromosomal markers, hybridization screening with libraries or flow-sorted chromosomal preparations specific to known chromosomes, and the like (Verma et al., Human Chromosomes: A Manual of Basic Techniques, Pergamon Press, New York N.Y., 1988). 
     To detect the presence of the splicing variants of genes predisposing an individual to prostate cancer, a test sample is prepared and analyzed for the presence or absence of such susceptibility alleles. Thus, the present invention provides methods to identify the expression of one of the nucleic acids of the present invention, or homolog thereof, in a test sample, using a nucleic acid probe or antibodies of the present invention. In particular, such methods comprise incubating a test sample with one or more of oligonucleotide probes of the present invention (as described above) and assaying for binding of the nucleic acid probes or antibodies to components within the test sample. 
     Conditions for incubating a nucleic acid probe or antibody with a test sample depend on the format employed in the assay, the detection methods used, and the type and nature of the probe used in the assay. One skilled in the art will recognize that any one of the commonly available hybridization or amplification formats can readily be adapted to employ the nucleic acid probes or antibodies of the present invention. Examples of such assays can be found in Chard, An Introduction to Radioimmunoassay and Related Techniques, Elsevier Science Publishers, Amsterdam, Netherlands, 1986; Bullock et al., Techniques in Immunocytochemistry, Academic Press, Orlando, Fla. Vol. 1, 1982, Vol. 2, 1983, Vol. 3, 1985; Tijssen, Practice and Theory of Immunoassays: Laboratory Techniques in Biochemistry and Molecular Biology, Elsevier Science Publishers, Amsterdam, Netherlands, 1985. 
     The test samples of the present invention include cells, protein or membrane extracts of cells, or biological fluids such as sputum, blood, serum, plasma, or urine. The test sample used in the above-described method will vary based on the assay format, nature of the detection method and the tissues, cells or extracts used as the sample to be assayed. Methods for preparing DNA extracts from any of the above samples are well known in the art and can be readily be adapted in order to obtain a sample which is compatible with the system utilized. 
     Gene Silencing 
     The phrase “gene silencing” refers to a process by which the expression of a specific gene product is lessened or attenuated. It is also used interchangeably with the term “gene knockdown.” Gene silencing can take place by a variety of pathways. Unless specified otherwise, as used herein, gene silencing refers to decreases in gene product expression that results from RNA interference (RNAi), a defined, though partially characterized pathway whereby small inhibitory RNA (siRNA) act in concert with host proteins (e.g. the RNA induced silencing complex, RISC) to degrade messenger RNA (mRNA) in a sequence-dependent fashion. The level of gene silencing can be measured by a variety of means, including, but not limited to, measurement of transcript levels by Northern Blot Analysis, B-DNA techniques, transcription-sensitive reporter constructs, expression profiling (e.g. DNA chips), and related technologies. Alternatively, the level of silencing can be measured by assessing the level of the protein encoded by a specific gene. This can be accomplished by performing a number of studies including Western Analysis, measuring the levels of expression of a reporter protein that has e.g. fluorescent properties (e.g. GFP) or enzymatic activity (e.g. alkaline phosphatases), or several other procedures. 
     The term “siRNA” refers to small inhibitory RNA duplexes that induce the RNA interference (RNAi) pathway. These molecules can vary in length (generally between 18-30 basepairs) and contain varying degrees of complementation to their target mRNA in the antisense strand. Some, but not all, siRNAs have unpaired overhanging bases on the 5′ or 3′ end of the sense strand and/or the antisense strand. The term “siRNA” includes duplexes of two separate strands, as well as single strands that can form hairpin structures comprising a duplex region. Designing a siRNA molecule that can specifically silence a certain gene is well known in the art, and can be routinely carried out using methods similar to what is disclosed in U.S. Pat. No. 8,008,474, which is incorporated herein by reference. siRNA can be routinely introduced to cells through conventional means such as transfection. 
     For targeted silencing of certain splicing variant, siRNA can be designed to target a specific exon that is only present in one variant. The mRNA of the variant that include this exon will be selectively silenced. Alternatively, siRNA can be designed to target a specific exon junction, which will only exist when certain splicing event occurs. In other words, siRNA can be designed to target the junction sequence of an exon immediately 5′ to the alternatively spliced exon and an exon that is immediately 3′ to the alternatively spliced exon. This particular junction sequence would only exist in a continuous polynucleotide sequence within an mRNA when the alternatively spliced exon is lacking.