KR101142889B1 - Screening Methods of Agents for Regulating the miRNAs and their targets which control Insulin Signaling Pathway - Google Patents

Abstract

The present invention relates to a method for screening a miRNA that regulates the insulin signal pathway and a substance that controls the action of the target gene, and specifically, USH or a target gene of miR-8 or miR-200 miRNA that promotes cell growth. A method for screening a substance that controls the action of FOG2. The inventors have discovered miR-8, a conserved miRNA that regulates body size by targeting u- shaped ( ush ) in Drosophila fat cells. In addition, it was confirmed that the target gene of miR-200, a human homologous gene of Drosophila miR-8 miRNA, is FOG2. Drosophila miR-8 and USH were also conserved in mammals and USH's human homologous gene, FOG2, was found to function directly in combination with the regulatory subunit of PI3K. Inhibition of miR-200 expression or expression of FOG2 in human cancer cell lines has been shown to reduce PI3K activity that promotes cell growth. Therefore, miR-200 and FOG2 can be usefully used for screening insulin signaling pathway regulators. .

Description

Screening methods of agents for regulating the miRNAs and their targets which control Insulin Signaling Pathway

The present invention relates to a method for screening substances that control the action of genes that regulate insulin signaling pathways.

Since an animal of abnormal size is culled because it is not suitable for survival, its body size is an important biological parameter for stable survival. So how is the body determined is a fundamental question in biology. Recent studies using insect model systems have reported that insulin signaling plays an important role in regulating body growth (Chen, C et. al ., 1996, Endocrinology 137, 846-856; Ikeya, T (2002, Curr Biol 12, 1293-1300; Rulifson, EJ et al ., 2002, Science 296, 1118-1120). Insulin (Insulin like peptide in Drosophila) binds to its receptor (IRS; chico in Drosophila) and triggers a phosphorylation cascade including insulin receptor substrate, phosphoinositide-3 kinase (PI3K) and Akt / PKB (Edgar, 2006) Mirth and Riddiford, 2007; Verdu et al . , 1999), the activated PI3K complex consists of an enzymatically active subunit (p110; dp110 in Drosophila) and a regulatory subunit (p85α; dp60 in Drosophila). Phosphorylated Akt (p-Akt) phosphorylates many proteins, including forkhead box O transcription factor (FOXO), which is involved in cell death, cell proliferation, metabolism, and life regulation (Gross, DN et al.). al ., 2008, Oncogene 27, 2320-2336). Once activated kinase cascade promotes cell growth and proliferation.

Growth of organisms can be achieved not only by cell-autonomous regulation but also by non-cell-autonomous regulation through circulation of growth hormones (Baker, J et. al ., 2004, Cell 116, 281-297; Edgar, BA 2006, Nat Rev Genet 7, 907-916; Lupu, F et al ., 2001, Dev Biol 229, 141-162; Mirth, CK, and Riddiford, LM 2007, Bioessays 29, 344-355; Velloso, CP 2008, Br J Pharmacol 154, 557-568). Recent studies have reported that in some endocrine organs, such as the insect's prothoracic gland and fat body, it regulates organism growth by adjusting development and nutritional conditions . ( Caldwell, PE et al ., 2005, Curr Biol 15, 1785-1795; Colombani, J et al ., 2005, Science 310, 667-670; Colombani, J et al ., 2003, Cell 114, 739-749; Mirth, C, et al ., 2005, Curr Biol 15, 1796-1807). However, the detailed mechanism of how body size is determined and controlled is still unknown.

microRNA (miRNA) is approximately 22 nt of untranslated RNA that acts as a post-transcriptional inhibitor through base binding with the 3 ′ untranslated region (UTR) site of mRNA (Bartel, 2004). The physiological function of each miRNA is still unknown. The study of miRNA function relies heavily on computer algorithms to predict target genes (Grun, D et al ., 2005, PLoS Comput Biol 1, e13; John, B et al ., 2004, PLoS Biol 2, e363; Kim, SK et al ., 2006, BMC Bioinformatics 7, 411; Kiriakidou, M et al ., 2004, Genes Dev 18, 1165-1178; Krek, A et al ., 2005, Nat Genet 37, 495-500; Lewis, BP et al ., 2005, Cell 120, 15-20; Stark, A et al ., 2003, PLoS Biol 1, E60), many false positive targets are predicted in spite of the usefulness of this target prediction program because regulation in the cell depends on the accessibility of the complementary sequence and the availability of the target message. Microarrays (Lim, LP et) to identify real targets al ., 2005, Nature 433, 769-773), proteomic analysis (Baek, D et al ., 2008, Nature 455, 64-71) and biochemical separation of miRNA-mRNA complexes (Beitzinger, M et. al ., 2007, RNA Biol 4, 76-84; Chi, SW et al ., 2009, Nature 460, 479-486; Easow, G et al ., 2007, RNA 13, 1198-1204; Karginov, FV et al ., 2007, Proc Natl Acad Sci USA 104, 19291-19296; Landthaler, M et al ., 2008, RNA 14, 2580-25915). Genetic approaches using model organisms may also be useful tools for studying the biological role of miRNAs ( J et. al ., 2003, Cell 113, 25-36; Giraldez, AJ et al., 2005, Science 308, 833-838; Ibanez-Ventoso, C et al ., 2008, PLoS ONE 3, e2818; Lee, RC et al ., 1993, Cell 75, 843-854; Rodriguez, A et al ., 2007, Science 316, 608-611; Thai, TH et al ., 2007, Science 316, 604-608; van Rooij, E et al ., 2007, Science 316, 575-579; Wightman, B et al ., 1993, Cell 75, 855-862; Zhao, Y et al ., 2007, Cell 129, 303-317). Despite these advances, it is not easy to identify the biological function of miRNA and its physiologically important targets.

Thus, the present inventors found that using Drosophila as a model system, the conserved miRNA miR-8 regulates the body population by targeting the Drosophila gene called u-shaped ( ush ) in fat cells. In addition, miR-8 and USH were also conserved in mammals, and it was found that FOG2, the human homologous gene of USH, functions in direct binding with the regulatory subunit of PI3K. In addition, it was confirmed that the target gene of miR-200, a human homologous gene of Drosophila miR-8 miRNA, is FOG2. Inhibition of miR-200 expression or expression of FOG2 in human cancer cell lines showed that PI3K activity promoting cell growth was decreased and cell growth was reduced. Therefore, miR-200 and FOG2 were used for the screening of insulin signaling pathway regulators. The present invention has been completed by confirming that it can be usefully used.

SUMMARY OF THE INVENTION

1) treating the test compound to a cell line expressing miR-200 family miRNA or miR-8 miRNA;

2) measuring the expression level or activity of FOG2 or USH protein in the cells of step 1) treated; And,

3) to provide a method for screening an insulin signaling regulatory substance comprising the step of selecting a test compound of which the expression or activity of the FOG2 or USH protein is changed in the cells of step 1) compared to the control.

Another object of the present invention

 1) treating the test compound to a cell line expressing FOG2 or USH;

2) measuring the expression level or activity of FOG2 or USH protein in the cells of step 1); And,

3) to provide a method for screening an insulin signaling regulatory substance comprising the step of selecting a test compound of which the expression or activity of the FOG2 or USH protein is changed in the cells of step 1) compared to the control.

Another object of the present invention

1) contacting the FOG2 protein with the p85 protein in the presence of the test compound;

2) measuring the degree of binding between the FOG2 protein and p85 protein in the above; And,

3) screening a test compound having a change in the degree of binding between the FOG2 protein and the p85 protein in the test tube of step 2) compared to a control which has not been treated with the test compound will be.

Another object of the present invention

1) contacting the USH protein with the dp60 protein in the presence of the test compound;

2) measuring the degree of binding between the USH protein and dp60 protein in the above; And,

3) screening for an insulin signaling regulatory substance comprising the step of selecting a test compound having a changed degree of binding between the USH protein and the dp60 (Drosophila p60) protein in the test tube of step 2) compared to the control which was not treated with the test compound. To provide a way.

Another object of the present invention

1) treating the test compound to a cell line expressing the FOG2 protein;

2) measuring the binding degree of FOG2 protein and p85 protein in the cells of step 1); And,

3) to provide a method for screening an insulin signaling regulatory substance comprising the step of selecting a test compound having a change in the binding degree of FOG2 protein and p85 protein in the cells of step 1) compared to a control that is not treated with the test compound. .

Another object of the present invention

1) treating the test compound to a cell line expressing the USH protein;

2) measuring the binding degree of USH protein and dp60 protein in the cells of step 1); And,

3) to provide a screening method of an insulin signaling regulatory substance comprising the step of selecting a test compound having a change in the degree of binding of USH protein and dp60 protein in the cells of step 1) compared to the control group not treated with the test compound. .

Another object of the present invention to provide a composition for regulating insulin signal transduction containing a substance that changes the expression or activity of FOG2 or USH protein as an active ingredient.

In order to achieve the above object, the present invention

1) treating the test compound to a cell line expressing miR-200 family miRNA or miR-8 miRNA;

2) measuring the expression level or activity of FOG2 or USH protein in the cells of step 1) treated; And,

3) provides a screening method of an insulin signaling regulatory substance comprising the step of selecting a test compound having a change in the expression or activity of FOG2 or USH protein in the cells of step 1) compared to the control.

In addition,

1) treating the test compound to a cell line expressing FOG2 or USH;

2) measuring the expression level or activity of FOG2 or USH protein in the cells of step 1); And,

3) provides a screening method of an insulin signaling regulatory substance comprising the step of selecting a test compound having a change in the expression or activity of FOG2 or USH protein in the cells of step 1) compared to the control.

In addition,

1) contacting the FOG2 protein with the p85 protein in the presence of the test compound;

2) measuring the degree of binding between the FOG2 protein and p85 protein in the above; And,

3) provides a screening method of an insulin signaling regulatory substance comprising the step of selecting a test compound of which the degree of binding between the FOG2 protein and the p85 protein is changed in the test tube of step 2) compared to a control which is not treated with the test compound. .

In addition,

1) contacting the USH protein with the dp60 protein in the presence of the test compound;

2) measuring the degree of binding between the USH protein and dp60 protein in the above; And,

3) screening for an insulin signaling regulatory substance comprising the step of selecting a test compound having a changed degree of binding between the USH protein and the dp60 (Drosophila p60) protein in the test tube of step 2) compared to the control which was not treated with the test compound. Provide a method.

In addition,

1) treating the test compound to a cell line expressing the FOG2 protein;

2) measuring the binding degree of FOG2 protein and p85 protein in the cells of step 1); And,

3) A method for screening an insulin signaling regulatory substance comprising the step of selecting a test compound having a change in the binding degree of FOG2 protein and p85 protein in the cells of step 1) compared to a control that is not treated with the test compound.

In addition,

1) treating the test compound to a cell line expressing the USH protein;

2) measuring the binding degree of USH protein and dp60 protein in the cells of step 1); And,

3) A method for screening an insulin signaling regulatory substance comprising the step of selecting a test compound having a change in the degree of binding of USH protein and dp60 protein in the cells of step 1) compared to a control that is not treated with the test compound.

 In addition, the present invention provides a composition for regulating insulin signal transduction containing a substance that changes the expression or activity of FOG2 or USH protein as an active ingredient.

We used Drosophila as a model system to discover miR-8, a conserved miRNA that regulates the body's sphere by targeting u-shaped (ush ) in adipocytes. In addition, it was confirmed that the target gene of miR-200, a human homologous gene of Drosophila miR-8 miRNA, is FOG2. Drosophila miR-8 and USH were also conserved in mammals and USH's human homologous gene, FOG2, was found to function directly in combination with the regulatory subunit of PI3K. Inhibition of miR-200 expression or expression of FOG2 in human cancer cell lines reduced PI3K activity that promotes cell growth. Therefore, miR-200 and FOG2 may be useful for screening insulin signaling regulators such as anticancer agents. Can be.

Hereinafter, the present invention will be described in detail.

In order to achieve the above object, the present invention

1) treating the test compound to a cell line expressing miR-200 family miRNA or miR-8 miRNA;

2) measuring the expression level or activity of FOG2 or USH protein in the cells of step 1) treated; And,

3) provides a screening method of an insulin signaling regulatory substance comprising the step of selecting a test compound having a change in the expression or activity of FOG2 or USH protein in the cells of step 1) compared to the control.

In addition,

1) treating the test compound to a cell line expressing FOG2 or USH;

2) measuring the expression level or activity of FOG2 or USH protein in the cells of step 1); And,

3) provides a screening method of an insulin signaling regulatory substance comprising the step of selecting a test compound having a change in the expression or activity of FOG2 or USH protein in the cells of step 1) compared to the control.

In addition,

1) contacting the FOG2 protein with the p85 protein in the presence of the test compound;

2) measuring the degree of binding between the FOG2 protein and p85 protein in the above; And,

3) provides a screening method of an insulin signaling regulatory substance comprising the step of selecting a test compound of which the degree of binding between the FOG2 protein and the p85 protein is changed in the test tube of step 2) compared to a control which is not treated with the test compound. .

In addition,

1) contacting the USH protein with the dp60 protein in the presence of the test compound;

2) measuring the degree of binding between the USH protein and dp60 protein in the above; And,

3) screening for an insulin signaling regulatory substance comprising the step of selecting a test compound having a changed degree of binding between the USH protein and the dp60 (Drosophila p60) protein in the test tube of step 2) compared to the control which was not treated with the test compound. Provide a method.

In addition,

1) treating the test compound to a cell line expressing the FOG2 protein;

2) measuring the binding degree of FOG2 protein and p85 protein in the cells of step 1); And,

3) A method for screening an insulin signaling regulatory substance comprising the step of selecting a test compound having a change in the binding degree of FOG2 protein and p85 protein in the cells of step 1) compared to a control that is not treated with the test compound.

In addition,

1) treating the test compound to a cell line expressing the USH protein;

2) measuring the binding degree of USH protein and dp60 protein in the cells of step 1); And,

3) A method for screening an insulin signaling regulatory substance comprising the step of selecting a test compound having a change in the degree of binding of USH protein and dp60 protein in the cells of step 1) compared to a control that is not treated with the test compound.

MiR-8 is known as the homologous homologue of the miR-200 family (see FIG. S1) that promotes the proliferation of human cancer cells (Ibanez-Ventoso et. al . , 2008). Thus, the present inventors attempted to reveal the biological function of miR-200 using Drosophila. It is known that the larvae become smaller when the larval stage of insects grows slower and / or when pupariation occurs faster, resulting in shorter larval growth stages (Colombani et. al . , 2003; Edgar, 2006; Mirth and Riddiford, 2007). We found that the body size and body weight of miR-8 null fruit flies, the body volume of miR-8 null larvae are quite small (see FIGS. 1A, 1B and 1C), and the small body sizes of these miR-8 null fruit flies are precocious. It was found that this is due to late growth during the larval stage rather than one solubilization, and not due to a decrease in cell size but due to a decrease in cell number (see FIGS. 1C and 1D).

In addition, the present inventors confirmed the activity of proteins related to insulin signaling in miR-8 null fruit flies to find out why miR-8 null fruit flies grow slowly, resulting in a significant decrease in insulin signaling in miR-8 null fruit flies. It was found that there is (see Fig. 2a and 2b).

The inventors looked at the spatial expression pattern of miR-8 in the larva, the miR-8 expression was the highest in the fat body of the larva (see Fig. 3a and 3b).

Recent studies have found that Drosophila fat is an important organ in regulating energy metabolism and growth (Colombani et. al . , 2005; Colombani et al . , 2003; Edgar, 2006; Leopold, P, and Perrimon, N 2007, Nature 450, 186-188). The present inventors confirmed that miR-8 expression of the larval fat body is important for Drosophila body regulation, and that the function of the miR-8 fat body differs from the neuron in a spatio-temporally specific manner (FIG. 4b and FIG. 5).

The present inventors sought to find out miR-8 target genes that regulate the body's structure to further investigate the function of miR-8 molecules. We assume that miR-8 is a highly conserved miRNA and that the target gene of miR-8 may also be involved in the cell proliferation phenotype because the human homolog of miR-8 promotes cell proliferation in human cells (see FIG. 6). It was. We used target prediction programs to select target gene candidates for Drosophila miR-8 and human miR-200 miRNAs (see FIG. 7A). Candidates selected for target were classified into two groups: Drosophila and human genes. Drosophila genes known as tumor suppressors or negative regulators of cell proliferation in at least one species in the target target candidate list ush, Lap1 , CG8445 , dbo , Lar , Ced -12, CG12333 , cbt , Scr , pan , pnut , Spri , Shc , Rassf And CG5 target8 , human orthologs of the target Drosophila genes FOG2 , SEPT7 , RIN2 , SHC1 , HOXB5 , KLF11, TCF7L1 , ERBB2IP , ELMO2 , BAP1 , WDR37 , KLHL20 , PTPRD , RASSF2 And VAC14 were selected as target genes. Drosophila gene u-shaped ( ush ) and its human homologous Friend of GATA 2 (FOG2, ZFPM2 ) was the most conserved of the genes predicted in these two species.

The present inventors selected seven gene pairs that respond to both miRNAs as targets of miR-8 and miR-200 family miRNAs among the selected target genes (see FIG. 7B), and then target genes of miR-8 miRNAs. Is ush and is a function for preventing neurodegeneration of miR-8 (Karres et. al . , 2007) as well as the features and spatial on body growth separation at the molecular level, and (see Fig. 8a), to determine that the USH inhibit body growth (see Fig. 8c), the miR-8 is ush in fat body It was found to be suppressed (see FIG. 9A). This shows that ush is more strongly inhibited in fat than other body parts. In addition, USH protein levels were observed to be significantly reduced than mRNA levels (see FIG. 9B), indicating that miR-8 inhibits USH expression through both destabilization and translation inhibition of USH mRNA.

In addition, the inventors have confirmed that miR-8 binds directly to the predicted target site of USH and mediates expression inhibition (see FIG. 9C), and the predicted target site for miR-8 is far from lineage.D. virilis AndD.grimshawiWas found in all Drosophilid species, includingushFound out that it is the true target of miR-8.

It has been found to control the organisms grow as cell autonomous (non-cell-autonomous) method bar (Britton, JS et - insulin signaling larval fat body from existing studies the ratio al ., 2002, Dev Cell 2, 239-249; Colombani et al . , 2005).

We have found that fruit flies appear to have a smaller body size (see FIG. 10A) and fewer wing cells (see FIG. 10B) than miR-8 null fruit flies when suppressing insulin signaling in Drosophila fat. Assuming that USH suppresses insulin signaling and inhibits body growth, we confirmed that the growth of fruit flies was inhibited when insulin signaling was suppressed in larval fat. It can be seen that delivery is specifically disrupted (see FIGS. 8C, 11A, 11B).

In addition, the present inventors have the further out the function of miR-8 in adipose tissue results, and by miR-8 is activated, the PI3K a cell autonomous (cell-autonomous) method promotes cell growth (see Figure 12), cell growth It was confirmed that the effect of miR-8 in can vary depending on the tissue expressed (see Fig. 13a, 13c, 13d).

In addition, the inventors confirmed that USH negatively regulates insulin signaling, and it was found that USH inhibits insulin signal transduction in a cellular autonomous manner in parallel with or above PI3K (FIGS. 14A, 14B and 14C). , FIG. 14D and FIG. 14E).

Excessive insulin signaling is known to reduce the levels of insulin receptor ( Inr ) and cytohesin Steppke ( step ) through negative feedback of FOXO (Fuss, B et al ., 2006, Nature 444, 945-948; Puig, O, and Tjian, R 2005, Genes Dev 19, 2435-2446). The inventors have confirmed that the reduction of insulin signaling when miR-8 is not expressed is recovered through knockdown of USH, and as a result, an increase in ush expression in the fat body of miR-8 null larvae is at least partly due to the lack of insulin signaling. It was found to affect (see Fig. 15).

Ush results of the miRNA target gene search mRNA had a predicted binding site of one miR-8, whereas ush 's mammalian ortholog , FOG2, had a predicted binding site of at least three miR-200 family miRNAs (see FIGS. 16A and 16B). . We believe that Drosophila miR-8 miRNA is ush Human miR-200 family miRNAs are also fog2 , just as they regulated expression by directly binding to the 3 'UTR of mRNA. It was confirmed that the binding directly to the 3 'UTR of the mRNA regulates the expression (see Figure 17).

FOG2 is expressed in the heart, brain, testes, liver, lungs and skeletal muscle (Holmes, M et. al ., 1999, J Biol Chem 274, 23491-23498; Lu, JR et al ., 1999, Mol Cell Biol 19, 4495-4502; Svensson, EC et al ., 1999, Proc Natl Acad Sci USA 96, 956-961; Tevosian, SG et al ., 1999, Proc Natl Acad Sci USA 96, 950-955). Despite the widespread expression of FOG2 in adult tissues, little is known about its function except in its role in embryonic heart development (Fossett, N, and Schulz, RA 2001, Trends). Cardiovasc Med 11, 185-190). On the other hand, miR-200 miRNA is known to be expressed in various adult organs including the pituitary gland, thyroid gland, islet, testes, prostate, ovary, breast and liver (Landgraf, P et. al ., 2007, Cell 129, 1401-1414. As a result of confirming the correlation between the expression of FOG2 protein and miR-200c cluster miRNA in human cell lines derived from different organs, in general, the expression levels of miR-200c cluster member and FOG2 had negative correlation ( 18).

In addition, we found that FOG2 was actually reduced by miR-200 miRNA (see FIGS. 19A, 19B and 18), and that Drosophila human miR-200 was conserved in the regulation of insulin signaling as in the case of miR-8 miRNA. It was confirmed that it has a role (see Figs. 19A, 19B and 19C). In addition, it was confirmed that FOG2 inhibited PI3K (see FIG. 20) and that phosphorylation of Akt was reduced by FOG2 (see FIG. 21A).

Akt inhibits FOXO activity, and analysis of the effect of miR-200 miRNA on the downstream transducer of Akt confirms that FOXO activity is decreased by miR-200 miRNA and FOXO activity is increased by FOG2 (FIG. 21b, 21c and 21d).

The present inventors confirmed that miR-200 specifically affects the insulin signaling pathway, and it was found that miR-200 specifically regulates PI3K-Akt-FOXO signaling (see FIGS. 21A and 21B). .

Stimulation of PI3K and AKT promotes cell proliferation and inhibits apoptosis (Pollak, 2008). Correspondingly, the introduction of miR-200 miRNA increased cell viability (see FIG. 23A). Introduction of inhibitors of miRNAs showed the opposite effect (see FIG. 23B). In addition, as a result of confirming the action mechanism of FOG2, it was found that FOG2 acts as a negative regulator of PI3K by inhibiting the formation of the IRS-1 / p85 / p110 complex (see FIG. 24).

Although FOG2 is known to be a nuclear transcription co-regulator, several studies have reported that FOG2 is also located in the cytoplasm (Bielinska, M et. al ., 2005, Endocrinology 146, 397M 3984; Clugston, RD et al ., 2008, Am J Physiol Lung Cell Mol Physiol 294, L665-675). This suggests that FOG2 is responsible for function in the cytoplasm (see FIGS. 25A and 25B).

As a result of experimenting whether FOG2 interacts with PI3K, it was confirmed that p85, which is a regulatory subunit of PI3K, directly binds to FOG2 (see FIGS. 26, 27, 28B and 28C).

In order to identify the domain of FOG2 that binds to p85, we have produced and identified several truncated mutations of FOG. As a result, the intermediate region of FOG2 (507-789 aa) mediates interaction with p85α (Fig. 27b), it was confirmed that the intermediate region plays an important role in inhibiting PI3K (see Figure 27c). The results suggest that FOG2 binds directly to p83 resulting in inhibition of PI3K activity. In particular, we found that Drosophila USH physically interacts with Drosophila p60 (dp60; Drosophila ortholog of p85α) when dp60 is co-expressed in USH and human HEK293T cells (see FIG. 29). This suggests that the mechanism of action of USH / FOG2 is preserved beyond species (see FIG. 30).

The cell line expressing the miR-200 family miRNA is a human cell line derived from heart, brain, testes, liver, lung and skeletal muscle tissue, and the cell line expressing the miR-8 miRNA is a Drosophila cell line derived from adipose tissue. Preferred but not limited to this.

Determination of the expression level of the FOG2 or USH protein is any one selected from the group consisting of Western blot, immunostaining, fluorescence staining and reporter assay, the binding of the FOG2 and p85 or USH and dp60 The measurement of p85 / p110 / IRS-1 complex formation is preferably performed by a co-immunoprecipitation method, but is not limited thereto.

The test compound of step 1) is any one selected from the group consisting of natural compounds, synthetic compounds, RNA, DNA, polypeptides, enzymes, proteins, ligands, antibodies, antigens, bacteria or fungi metabolites and bioactive molecules, It is preferable to use substances whose antitumor effect is not known to those skilled in the art.

The activity of the FOG2 or USH protein is

Iii) measuring the activity of the p85 / p110 complex;

Ii) measuring the expression level of FOXO mRNA or protein; And,

Ⅲ can be measured by any one method selected from the group consisting of a method for measuring cell growth and proliferation, the activity of the p85 / p110 complex ( PI3K) can determine the degree of phosphorylation of AKT protein or the measurement of kinase activity of PI3K It may be measured by measuring, but is not limited thereto.

The binding between the proteins can be measured by any one method selected from the group consisting of SPR, protein chip, gel shift assay, immunoprecipitation, co-immunoassay, fluorescence immunoassay and radioimmunoassay. However, the present invention is not limited thereto, and may be measured through any method for measuring binding of proteins commonly used in the art.

 In addition, the present invention provides a composition for regulating insulin signal transduction containing a substance that changes the expression or activity of FOG2 or USH protein as an active ingredient.

The substance that changes the expression of the FOG2 or USH protein may be an expression inhibitor of FOG2 or USH protein, the expression inhibitor of the FOG2 or USH protein is fog2 or ush It is preferably an antisense nucleotide or small interfering RNA (siRNA) that complementarily binds to the mRNA of the gene. In addition, the substance that changes the activity of the FOG2 or USH protein may be an activity inhibitor of the FOG2 or USH protein, the activity inhibitor of the FOG2 or USH protein is a cytotoxic compound, peptide, It is preferably any one selected from the group consisting of peptide mimetics and antibodies, but is not limited thereto.

The siRNA is an antisense sequence that complementarily binds to the sense sequence of 15 to 30 mers selected from the base sequence of the mRNA of the gene encoding FOG2 or USH (SEQ ID NO: 42 or SEQ ID NO: 15). In this case, the sense sequence is not particularly limited thereto, but is preferably composed of 19-27 bases, and more preferably has a nucleotide sequence set forth in SEQ ID NO: 119 in the case of FOG2 protein expression inhibitor.

The antisense nucleotides, as defined in Watson-click base pairs, bind (hybridize) the complementary sequencing of DNA, immature-mRNA or mature mRNA to hinder the flow of genetic information as a protein in DNA. The nature of antisense nucleotides specific to the target sequence makes them exceptionally multifunctional. Since antisense nucleotides are long chains of monomeric units they can be easily synthesized for the target RNA sequence. Many recent studies have demonstrated the usefulness of antisense nucleotides as biochemical means for studying target proteins (Rothenberg et al. al ., J. Natl . Cancer Inst . , 81: 1539-1544, 1999). The use of antisense nucleotides can be considered as a new type of inhibitor because of recent advances in nucleotide synthesis and in the field of nucleotide synthesis that exhibit improved cell adsorption, target binding affinity and nuclease resistance.

Peptide Minetics are peptides or non-peptides that inhibit the binding domain of FOG2 or USH protein and inhibit the activity of FOG2 or USH protein. Major residues of non-hydrolyzable peptide analogs include β-turn dipeptide cores (Nagai et. al . 1985, Tetrahedron Lett 26: 647), keto-methylene pseudopeptides (Ewenson et al . 1986, J Med Chem 29: 295; And Ewenson et al . 1985, in Peptides: Structure and Function (Proceedings of the 9th American Peptide Symposium) Pierce Chemical Co. Rockland, IL), Azepine (Huffman et) al . 1988, in Peptides: Chemistry and Biology, GR Marshall ed., ESCOM Publisher: Leiden, Netherlands), benzodiazepines (Freidinger et al . 1988, in Peptides; Chemistry and Biology, GR Marshall ed., ESCOM Publisher: Leiden, Netherlands), β-aminoalcohol (Gordon et al . 1985, Biochem Biophys Res Commun 126: 419) and substituted gamma lactam ring (Garvey et al . , 1988 in Peptides: Chemistry and Biology, GR Marshell ed., ESCOM Publisher: Leiden, Netherlands.

The composition of the present invention comprises 0.0001 to 50% by weight of the active ingredient based on the total weight of the composition.

The composition of the present invention may contain one or more active ingredients exhibiting the same or similar function in addition to the substance for changing the expression or activity of the FOG2 or USH protein.

The composition of the present invention may further comprise at least one pharmaceutically acceptable carrier in addition to the above-described effective ingredients for administration. Pharmaceutically acceptable carriers may be used in combination with saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, liposomes, and one or more of these components, as needed. Other conventional additives such as buffer, bacteriostatic and the like can be added. It can also be formulated into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions, etc., with the addition of diluents, dispersants, surfactants, binders and lubricants additionally, and specifically for organs. To act and organ specific antibodies or other ligands can be used in combination with the carrier. It is better suited to formulate according to the respective disease or component by an appropriate method of the technical need or as disclosed in Remington's Pharmaceutical Science (Recent Edition, Mack Publishing Company, Easton PA). Can be.

Nucleotides or nucleic acids used in the present invention can be prepared for the purpose of oral, topical, parenteral, intranasal, intravenous, intramuscular, subcutaneous, intraocular, transdermal and the like. Preferably, the nucleic acid or vector is used in injectable form. Thus, in particular, the area to be treated may be mixed with any pharmaceutically acceptable vehicle for injectable compositions for direct infusion. The composition of the present invention may in particular comprise an isotonic sterile solution or a lyophilized composition which allows the composition of an injectable solution upon the addition of dry, in particular sterile water or a suitable physiological saline solution. Direct injection of nucleic acid into a patient's tumor is advantageous because it allows the treatment efficiency to be focused on the infected tissue. The dosage of nucleic acid used can be adjusted by various parameters, in particular by gene, vector, mode of administration used, disease in question or alternatively desired duration of treatment. In addition, the range varies depending on the weight, age, sex, health status, diet, administration time, administration method, excretion rate and severity of the patient. The daily dosage is about 0.01 to 12.5 mg / kg, preferably 1.25 to 2.5 mg / kg, preferably administered once to several times a day.

The composition for controlling insulin signaling of the present invention is useful for molecular biological studies of the insulin signaling pathway and for the prevention or treatment of diseases caused by abnormally functioning insulin signaling pathways such as cancer, diabetes and obesity. Could be used.

Hereinafter, the present invention will be described in detail by the following examples.

However, the following examples are illustrative of the present invention, and the contents of the present invention are not limited by the following examples.

Example 1 . Transgenic Drosophila and Drosophila Cultures

The characteristics, sources and references of the transgenic Drosophila used in the present invention are shown in Table 1. The fruit flies were incubated in standard Drosophila medium at 25 ° C. and used for the experiment.

Drosophila Characteristic Where to get References mir-8 △ 2 miR-8 null Drosophila Steve Cohen Karres et al . , 2007 mir-8 gal4 UAS - in combination with GFP expressing GFP by regulating the mir -8 enhancer Kyoto Stock Center Karres et al . , 2007 ush1513 Drosophila containing ush1513 low-order gene (hypomorph) that contains mutations in the promoter region and expresses ush at low levels Pat simpson Cubadda, Y et al ., 1997 Cg gal4 Gal4 expression in fat body and anterior lymph gland Young Joon Kim Takata et al ., 2004 ppl gal4 Adipose-specific expression of Gal4 / attenuated in salivary glands M. Pankratz Zinke et al . , 1999

Example 2 . Cell line culture

The cell lines of Table 2 purchased from the American Cell Line Bank (ATCC) were incubated at 37 ° C., 5% CO ₂ conditions.

Cell lines cultured above were removed from 75-cell culture flasks by trypsin-ethylenediamine tetraacetic acid (Invitrogen, USA) treated with trypsin-EDTA, and then inactivated trypsin by adding medium containing serum, followed by centrifugation. Precipitated. After removing the supernatant, the cells were suspended by adding culture medium for each cell line. Living cells were stained with trypan blue dye exclusion test and counted using a hemocytometer, followed by subculture at 5 × 10 ⁵ cells / flask in a 100 mm dish.

Cell line badge HepG2 RPMI-1640 Huh7 AsPC1 PANC1 SW480 MCF7 MDA MB 231 U2OS HeLa Hep3B IMDM HCT116 HCT116p53KD

* IMDM (Iscove's Modified Dulbecco's Medium, Gibco, USA): Contains 10% FBS (Gibco)

RPMI-1640 (Gibco): 10% FBS

Example 3 . Small Body Phenotype of the mir -8 Mutant Drosophila

Transduction of human miRNA miR-200 family (miR-200a, miR-200b, miR-200c, miR-141, and miR-429) into human cell lines has been known to promote cell growth (Park, SY). et al ., 2009, Nat Struct Mol Biol 16, 23-29). We treated the MCF-7 cell line with nothing, siRNA against GFP (siGFP, SEQ ID NO: 5'-UGAAUUAGAUGGCGAUGUU-3 ', Samchully), miR-200 family [miR-141 (has-miR-141, sequence) No. 2: 5'-UAACACUGUCUGGUAAAGAUGG-3 '), miR-200b (has-miR-200b, SEQ ID NO: 3: 5'-UAAUACUGCCUGGUAAUGAUGA-3'), miR-429 (has-miR-429, SEQ ID NO: 4: 5 '-UAAUACUGUCUGGUAAAACCGU-3')] and miR-29b (known to inhibit cell proliferation) in MCF-7 cell lines transduced and counted on days 1, 4, 5, 6, 7 and 8 As a result of measuring the cell proliferation, when the miR-200 family was treated, nothing was treated, or the cell proliferation was increased than the siGFP treatment, and the miR-29b treatment almost stopped cell proliferation from the 5th day. , miR-200 family once again confirmed to promote cell proliferation in human cells (FIG. 6).

The miR-200 family has increased expression in cancer cells, including uterine cancer (Iorio, MV et al ., 2007, Cancer Res 67, 8699-8707; Nam, EJ et al ., 2008, Clin Cancer Res 14, 2690-2695), highly conserved in bilaterian animals, and miR-8 is known as the homologous gene of miR-200 in Drosophila melanogaster (Ibanez-Ventoso et. al . , 2008). Thus, the present inventors used fruit flies to reveal the biological function of miR-200. The present inventors first miR-8 null (null) Drosophila ^{[mir -8 △ 2 (Karres,} JS et al ., 2007, Cell 131, 136-145) observed that the body size and body weight were significantly smaller compared to wild-type Drosophila (FIGS. 1A and 1B). The body weight of the fruit flies was measured after ice anesthesia of 5 to 3 day old fruit flies after the incubation of each group. In addition, miR-8 null fruit flies are known to have high frequency of apoptosis in the brain and high frequency of malformed legs and wings (Karres et al. al . , 2007).

Determination of the final body size of insects during larvae is similar to that of growth in human adolescence (Edgar, 2006; Mirth and Riddiford, 2007). It is known that the larvae become smaller when the larval stage of insects grows slower and / or when pupariation occurs faster, resulting in shorter larval growth stages (Colombani et. al . , 2003; Edgar, 2006; Mirth and Riddiford, 2007). We compared the miR-8 null larvae at 100 hours of hatching with wild-type larvae, with a significantly smaller body volume (FIG. 1C), with each group recovering eggs after 5 hours of spawning, and pupae every 12 hours. As a result of counting the number of adults, miR-8 null fruit flies were not significantly different from wild type fruit flies (Fig. 1c), but allegory (departure from emergence, pupae and adult) was delayed by about 12 hours. It was confirmed that (Fig. 1c). The small size of the miR-8 null fruit flies was due to late growth during the larval stage, rather than premature solubilization.

Lack of food intake is known to be accompanied by premature or delayed solubility with reduced signs of eating (Layalle, S et. al ., 2008, Dev Cell 15, 568-577; Mirth et al . , 2005). However, fruit flies are known whose expression is decreased in starvation conditions, insulin-like peptide (Drosophila insulin-like peptides, Dilps) (Colombani et al . , 2003; Ikeya et al . , 2002), the expression was not reduced in miR-8 null larvae. Influence of Dilps expression and solubilization timing in miR-8 null larvae means that the small populations of miR-8 null fly flies are not due to reduced feeding.

Thus, the inventors have taken 8 to 10 wings of miR-8 null fruit flies and wild type fruit flies to determine whether the small body phenotype is due to a decrease in cell size and / or cell number. The relative size of the cells was calculated by counting the number of wing hairs, and the relative size of the cells was measured by dividing the whole wing pack cell region into the cell pixel region. As a result, the miR-8 null fruit flies did not show a significant difference in the size of the wing cells as compared to the wild type fruit flies, but the number of wing cells was smaller (FIG. 1D). From the above results, it can be seen that the reason for reducing the growth of the terminal tissue of miR-8 null Drosophila is not the decrease of the cell size but the decrease of the number of cells.

Example 4 Decreased Insulin Signaling in miR- 8 Null Drosophila

To determine why miR-8 null fruit flies grow slowly, we identified the activity and gene expression of proteins involved in insulin signaling in miR-8 null fruit flies. First, the levels of activated Akt (p-Akt) were confirmed by Western blot using anti-phosphorylated Akt specific antibodies (FIG. 2A). Specifically, 300 μl of lysis buffer solution (1% Triton X-100, 50 mM Tris pH7.4, 500 mM NaCl, 7.5 mM MgCl2, 0.2) was used in wild-type and miR-8 null Drosophila, respectively. After homogenization using a known method using mM EDTA, 1 mM NaVO 4, 50 mM β-glycerophosphate, 1 mM DTT, 25% glycerol ( Lee, SB et al., 2007, EMBO Rep 8 , 360-365), RIPA buffer (25 mM Tris, pH 7.4, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, each group separated by using a 0.1% SDS) Proteins were separated using 6-10% SDS-PAGE. The isolated protein was transferred to a PVDF membrane using a semi-dry transfer method, incubated in a 2% BSA solution, blocked, and then subjected to anti-phosphorylation-Akt (P-Akt) antibody [1: 1,000, ( # 9271S, Ser473) Cell Signaling Technology, USA] in 2% BSA solution containing. Then, incubated in a 2% BSA solution containing a peroxidase-attached anti-mouse secondary antibody (SantaCruz, USA), and then using a chemiluminescence substrate (Chemiluminescence, Roche, USA) using immunochemistry Was developed on the X-ray film to check the band. As a result, it was confirmed that the level of p-Akt was reduced in miR-8 null Drosophila, it was found that the Akt signal transduction process is not properly performed without miR-8 (Fig. 2a).

Activated p-Akt is known to phosphorylate and inactivate FOXO. Phosphorylation of FOXO prevents nuclear localization, which reduces transcription of the FOXO target gene. The present inventors investigated the expression level of 4EBP, a FOXO target gene, through 4EBP gene Q-PCR in miR-8 null larvae. First, the whole RNA was extracted from the wild larvae and miR-8 null Drosophila larvae (96 hours after incubation) using Trizol reagent. After filling up to 50 μl, RT-PCR reaction was performed in a SuperScript First-Strand Synthesis System (Invitrogen, USA). CDNA was synthesized by reacting at 65 ° C. for 6 minutes, 4 ° C. for 5 minutes, 42 ° C. for 60 minutes, 95 ° C. for 5 minutes, and 4 ° C. for 5 minutes. Primer pairs of 4 ebp sense primers (SEQ ID NO: 5'-ATGCAGCAACTGCCAAATC-3 ') and 4 ebp antisense primers (SEQ ID NO 6: 5'-CCGAGAGAGAAAACAAGGTGG--3') and the ABI SYBR PCR using the cDNA as a template Quantitative PCR for 4ebp mRNA was performed in the ABI Prism 7900 Sequence Detection System (Applied Biosystems, USA) using a master mix ( Applied Biosystems, USA). At this time, the rp49 gene was PCR together as a quantitative control using primer pairs of rp49 sense primer (SEQ ID NO: 5'-AGGGTATCGACAACAGAGTG-3 ') and rp49 antisense primer (SEQ ID NO: 8: 5'-CACCAGGAACTTCTTGAATC-3'). . 4 ebp mRNA expression was converted to a relative value according to the amount of rp49 mRNA, the Ct (comparative cycle threshold) value was analyzed according to the manual (User Bulletin 2, Applied Biosystems, USA). Repeating the experiment three times, and then, are shown graphically on the average (Fig. 2b). From this, it can be seen that insulin signaling is significantly reduced in miR-8 null Drosophila.

Example 5 . Identification of miR- 8 expression level in Drosophila tissue

Cohen et al. Reported the expression of miR-8 in Drosophila brain, wing discs and leg discs (Karres et al. al . , 2007). We describe the spatial expression pattern of miR-8 in larvae by the mir- 8 enhancer trap GAL4 strain Drosophila ( mir- 8 gal4 ) (Karres et al. al . , 2007) of the third body larvae (96 hours after spawning) fat body, cuticles and brain tissues by immunostaining by known methods (Lee, Y et al ., 2005, Nat Genet 37, 305-310), GFP expression was the highest in the fat body (Fig. 3a). In addition, the total RNA was isolated from the total larvae, fat body, intestine, brain and cuticles and the miR-8 expression level was confirmed through Northern blot. Specifically, the whole RNA was extracted using Trizol reagent (Invitrogen, USA) from fat, intestine, brain, and cuticle dissected by dissecting the entire miR-8 Gal4 Drosophila and the miR-8 Gal4 Drosophila, and then total RNA of each tissue. 5.4 μg was run on a 12.5% urea-polyacrylamide gel. The unfolded RNA was delivered with current through a Zeta probe GT membrane (Biorad, USA). Oligonucleotides corresponding to miR-8 were end-labeled with [γ- ³² ATP] and used as probes of the northern blot. The membrane treated with the probe was exposed to a phosphorylated imaging plate (fugi film, Japan), and then confirmed using a BAS-2500 system (Fugi). As a result, the miR-8 expression was the highest in the fat body of the larvae (FIG. 3B).

Example 6 fat body of miR- 8 ( fat body ) -specific expression of Drosophila body size

Recent studies have found that Drosophila fat is an important organ in regulating energy metabolism and growth (Colombani et. al . , 2005; Colombani et al . , 2003; Edgar, 2006; Leopold and Perrimon, 2007). The inventors hypothesized that the miR-8 expression level of the larval fat body is important for the regulation of Drosophila body size, and to observe the weight of Drosophila when the miR-8 null Drosophila specifically expressed miR-8. The following experiment was performed.

<6-1> miR -8 and miR -200 miRNA  Preparation of Expression Vectors

The inventors performed cloning as follows to prepare transgenic Drosophila expressing miR-8 miRNA and miR-200c miRNA cluster. First, in order to obtain "genome region of miR-8 miRNA" described in SEQ ID NO: 9 (5'-TAATACTGTCAGGTAAAGATGTC-3 '), genomic DNA is extracted from Drosophila using Trizol reagent, and then the genomic DNA (3 ~ PCR was performed using primer pairs of SEQ ID NO: 10 (5'-TCACAAAGAATTCCTTATCGCCA-3 ') and SEQ ID NO: 11 (5'-GCTCTAGAATTTTCAGCCGCTTGTCTTCGC-3') and PCR Mastermix (Promega, USA) It was. PCR products of miR-8 generated after the completion of the reaction were digested with restriction enzymes of Eco RI, and then inserted into pUAS vectors digested with the same restriction enzymes to prepare a UAS - mir- 8 vector.

In addition, the inventors of the miR-200c To prepare a transgenic Drosophila expressing human miR-200c cluster comprising mir - 141 , cloning was performed as follows. First, genomic DNA was extracted from the cell line HeLa cell line expressing "miR-200c cluster" described in SEQ ID NO: 12 using Trizol reagent, and then the genomic DNA (3-5 µg) was used as a template. MiR-200c using primer pairs of (5'-CGGAATTCACGGTCGAGGGGCTTCGGAG-3 ') and SEQ ID NO: 14 (5'-GCTCTAGACATGCTCCCAAGGCGGGAAGAC-3') and PCR Mastermix (Promega, USA) And PCR products for polynucleotides encoding human miR-200c cluster comprising mir - 141 . The PCR product was digested with restriction enzymes of Eco RI and Xba I, and then inserted into a pUAS vector digested with the same restriction enzyme to be UAS - human miR-200c. Cluster vectors were prepared.

<6-2> Fat body  Specifically miR Expressing -8 miR Preparation of the -8 Defective Drosophila

Table 1 mir -8 ^{△ 2} and Cg It crossed the gal4 miR-8 the deficit Cg gal4 fruit fly mir8 CgG4 fruit flies were selected. Further, in Table 1 of the mir -8 ^{△ 2} and ppl ppl missing miR-8 by crossing gal4 We were selected gal4 mir8 pplG4 of Drosophila fruit flies. Each mir -8 ^{△ 2} , Cg gal4 , ppl gal4 The fruit flies are mated with the double balancer fruit flies and mir8 is the standard breeding method. CgG4 and mir8 pplG4 Drosophila After that, mir8 to the UAS - mir- 8 vector prepared in Example 1-1. CgG4 And mir8 pplG4 is transformed through a known method of P-element-mediated germ line transformation to mir8 CgG4 > mir8 And mir8 pplG4 > mir8 Drosophila was obtained (Rubin GM et al ., 1982, 357 Science 218: 348-353). In addition, the UAS - human miR-200c prepared in Example 1-1 Mir8 cluster vector CgG4 And mir8 pplG4 was transformed in the same manner as above to mir8 CgG4 > 200c and mir8 pplG4 > 200c Drosophila were obtained.

<6-3> miR -8 defect In Drosophila Fat body  Specific miR Of Eight-8 Expression on Drosophila Growth

The present inventors fat body - Cg having a specific Gal4 driver of Cg and ppl gal4 , mir8 CgG4 , mir8 CgG4> mir8 and mir8 CgG4 > 200c or , ppl gal4 , mir8 pplG4 , mir8 pplG4 > mir8 and mir8 pplG4> 200c body weight was measured in the same manner as in Example 3, when miR-8 null Drosophila specifically expressed miR-8 in the body ( mir8 CgG4 > mir8 and mir8 pplG4 ), wild-type fruit fly ( Cg gal4 and ppl gal4 ) was confirmed to increase in weight to a similar degree (Fig. 4a). This supports the miR-8 specific function of fat and suggests that miR-8 of fat is important for overall body growth. In addition, when human miR-200 cluster was expressed in Drosophila fat body ( mir8 CgG4 > mir8 and mir8 pplG4> mir8 ) also increased the weight of fruit flies (FIG. 4A). Human miR-200 family miRNAs present in both chromosome clusters are highly homologous to miR-8 (FIG. 5). The fact that human miR-200c clusters effectively replace the role of miR-8 is because human miRNAs have been processed through the Drosophila miRNA processing instrument, which means that they share conserved biological functions.

<6-4> miR -8 defect In Drosophila  Neuron specific miR Of Eight-8 Expression on Drosophila Growth

As mentioned above, miR-8 is expressed in the central nervous system and is known to prevent neurodegeneration (Karres et. al . , 2007). miR-8 nerve in Table 1 in order to indirectly control the feeding behavior in cells involved in the regulation of Learn how organisms grow mir -8 ^{△ 2} and elav elav missing miR-8 by crossing gal4 gal4 fruit fly mir8 the elavG4 flies were selected. Each mir -8 ^{△ 2} and elav gal4 Drosophila is bred with a double balancer Drosophila, respectively, using the standard breeding method mir -8 ^{△ 2} and elav I made a fruit fly with gal4 . The UAS - mir- 8 vector prepared in Example 1-1 was transformed into mir8 elavG4 by the same method as described above. elavG4 > mir8 fruit flies were obtained. mir8 elavG4 and mir8 As a result of measuring the weight of elavG4 > mir8, the weight of Drosophila was similar regardless of whether miR-8 was expressed in neurons (FIG. 4B). This means that the function in the fat body of miR-8 is SPATIO-temporally specific to neurons.

Example 7 Target gene selection of miR -8 and miR -200

The present inventors sought to find out miR-8 target genes that regulate the body's structure to further investigate the function of miR-8 molecules. We hypothesized that miR-8 is a highly conserved miRNA, and that the human homologous gene of miR-8 promotes cell proliferation in human cells (FIG. 6A), so that the target gene of miR-8 may also be involved in the cell proliferation phenotype. . We used five target prediction programs (FIG. 7A) to select target gene candidates for Drosophila miR-8 and human miR-200 miRNA: miRanda (John et al . , 2004), miTarget (Kim et al . , 2006), microT (Kiriakidou et al . , 2004), PicTar (Krek et al . , 2005), and Target Scan (Lewis et al . , 2005). Candidates selected above were classified into two groups: Drosophila and human genes, Ensembl Orthologous data (//www.ensembl.org/) and NCBI Homologene data (//www.ncbi.nlm.nih.gov/Homologene/) And Inparanoid data (//inparanoid.sbc.su.se/) were used to identify species homologous gene pairs in the two groups (FIG. 6B). The inventors have identified Drosophila genes known as tumor suppressors or negative regulators of cell proliferation in at least one species in the target candidate list ush , Lap1 , CG8445 , dbo , Lar , Ced-12, CG12333 , cbt , Scr , pan , pnut , Spri , Shc , Rassf And CG5608 , the human orthologs of each Drosophila gene FOG2 , SEPT7 , RIN2 , SHC1 , HOXB5 , KLF11 , TCF7L1 , ERBB2IP , ELMO2 , BAP1 , WDR37, KLHL20 , PTPRD , RASSF2 And VAC14 were selected as target genes. Drosophila gene u-shaped (ush ) and its human homologous Friend of GATA 2 ( FOG2 , ZFPM2 ) was the most conserved of the predicted genes in these two species.

Example 8 . Target gene selection of miR- 8 plays an important role in the regulation of body growth

<8-1> miR -8 and / or miR -200 miRNA Preparation of Reporter Gene Expression Vectors for Target Genes

The inventors inserted a luciferase gene downstream of the 3 'UTR of each target gene including the miRNA target site to confirm that the target gene selected in Example 7 was actually a target of miR-8 and miR-200 family miRNAs. Cloning was carried out. Specifically, in order to obtain cDNA of each target gene, after extracting the total RNA from Drosophila S2 cell line or human Hela cell line using RNeasy mini kit (QIAGEN, USA), 1 μg total RNA and 1 μl Oligo-dT (Invitrogen , USA) was filled with 50 μl of distilled water, and then placed in a SuperScript First-Strand Synthesis System (Invitrogen, USA) for RT-PCR reaction. CDNA was synthesized by reacting at 65 ° C. for 6 minutes, 4 ° C. for 5 minutes, 42 ° C. for 60 minutes, 95 ° C. for 5 minutes, and 4 ° C. for 5 minutes. The cDNA (3 ~ 5 ㎍) template synthesis, ush , Lap1 , CG8445 , dbo among the target genes selected in Example 7 using the primer pairs and PCR Mastermix (Promega, USA) for each gene of Table 3 , Lar , Ced -12, CG12333 , cbt , FOG2 , SEPT7 , RIN2 , SHC1, HOXB5 , LF11 , TCF7L1 , ERBB2IP , ELMO2 , BAP1 , WDR37 , KLHL20 , PTPRD , RASSF2 And PCR for 3 'UTR of VAC14 was performed respectively. At this time, it was carried out with PCRP, on atro contrast to the Drosophila gene. After completion of the reaction, PCR products for the 3 'UTR region of each gene generated were digested with the following restriction enzymes:

ush , atro , CG 8445, cbt , lap 1, Ced - 12, CG 12333, dbo and lar use Sac I and Bam HI restriction enzymes;

FOG 2 , SEPT 7 , HOXB 5, KLF 11 , ERBB 2IP, ELMO 2, BAP 1 , PTPRD using Xba I and Eco RI restriction enzymes;

RIN 2, SHC 1 , WDR 37 , KLHL 20 , RASSF 2 , VAC 14 use Xba I restriction enzymes; And,

TCF 7l1 uses Eco RI and Eco RV restriction enzymes.

A reporter plasmid having a miR-8 or miR-200 family target sequence in the 3 'UTR sequence of the luciferase gene was prepared by inserting into a pAC5.1 luciferase vector (a kind gift of Elisa Izaurralde) digested with the same restriction enzyme. This is shown in Table 4.

origin gene SEQ ID NO: Primer sequence SEQ ID NO: Sense primer SEQ ID NO: Antisense primer Drosophila ush SEQ ID NO: 15 SEQ ID NO: 16 5'-ATCGATGAGCTCGAAAGCCAGCTGCGGTGGGAG-3 ' SEQ ID NO: 17 5'-ACGCGGATCCGGCAGCACTCAAATCGTTCGTTG-3 ' Drosophila atro SEQ ID NO: 18 SEQ ID NO: 19 5'-ATCGATGAGCTCGTGGACAGACAGCAATTGTAGAGAGC-3 ' SEQ ID NO: 20 5'-ACGCGGATCCGCAGAGAAGGTTTTCTGGGATTTGG-3 ' Drosophila CG8445 SEQ ID NO: 21 SEQ ID NO: 22 5'-ATCGATGAGCTCCCAGACGACCAGCATGTACATG-3 ' SEQ ID NO: 23 5'-ACGCGGATCCCAATAACGTTAGGATTCACCCGATT-3 ' Drosophila cbt SEQ ID NO: 24 SEQ ID NO: 25 5'-ATCGATGAGCTCGAAGAGCAATGCGAGCAGCATC-3 ' SEQ ID NO: 26 5'-ACGCGGATCCGATTGGAATCGTGCGATGATCC-3 ' Drosophila Lap1 SEQ ID NO: 27 SEQ ID NO: 28 5'-GAGCTCTTATTGTCCAATCGATCGAAGTGTC-3 ' SEQ ID NO: 29 5'-GGATCCGCGTGTGTTCTTCAATAATGCA-3 ' Drosophila Ced -12 SEQ ID NO: 30 SEQ ID NO: 31 5'-GAGCTCCATAACGAGCACAATTACTTCACG-3 ' SEQ ID NO: 32 5'-GGATCCCGTCACCAGTTTGTTCATTTTG-3 ' Drosophila CG12333 SEQ ID NO: 33 SEQ ID NO: 34 5'-GAGCTCTCTGCGAAATCCCAAACATCT-3 ' SEQ ID NO: 35 5'-GGATCCCGCTATGCCACGACTATTTGG-3 ' Drosophila dbo SEQ ID NO: 36 SEQ ID NO: 37 5'-GAGCTCTGTTTATTTGGTAGAGCCTAAAGCG-3 ' SEQ ID NO: 38 5'-GGATCCCCACCGGCAACATTTAGATAT-3 ' Drosophila Lar SEQ ID NO: 39 SEQ ID NO: 40 5'-GAGCTCCAATGGGATTGCCAGGTCCAC-3 ' SEQ ID NO: 41 5'-GGATCCTCCCCCAAAGAATCGTAATGG-3 ' human FOG2 SEQ ID NO: 42 SEQ ID NO: 43 5'-GCTCTAGACAGTCACCTTTGGTATCAGTGTTTAG-3 ' SEQ ID NO: 44 5'-CGGAATTCCATACTTGCAGCATTGAGTTTAGGG-3 ' human SEPT7 SEQ ID NO: 45 SEQ ID NO: 46 5'-GCTCTAGAGACCACCAGTTAACGTATTAGTTGCC-3 ' SEQ ID NO: 47 5'-CGGAATTCGAGGCCATGATTCCCATTTCTAGT-3 ' human RIN2 SEQ ID NO: 48 SEQ ID NO: 49 5'-GCTCTAGACTTCCCAGTGGTGCATCCAAAGG-3 ' SEQ ID NO: 50 5'-CATAAGTTTAATTTCACTGCAGGTTCTG-3 ' human SHC1 SEQ ID NO: 51 SEQ ID NO: 52 5'-GCTCTAGACTAGCGCTCTCTTCCAGAAGATGC-3 ' SEQ ID NO: 53 5'-GTACTCAGGAACTGCAGGTTTTGC-3 ' human HOXB5 SEQ ID NO: 54 SEQ ID NO: 55 5'-GCTCTAGACAGAGGAGCCCAGCGGCCCA-3 ' SEQ ID NO: 56 5'-CGGAATTCCACTGAGACAGGCCCCGCAAAGACA-3 ' human KLF11 SEQ ID NO: 57 SEQ ID NO: 58 5'-TCTAGAGGTCCATTAGGACATCACTC-3 ' SEQ ID NO: 59 5'-GAATTCCTGCCCAGGCAATAAAGTG-3 ' human TCF7L1 SEQ ID NO: 60 SEQ ID NO: 61 5'-GAATTCGACCCCTGCAGGCTGTCAC-3 ' SEQ ID NO: 62 5'-GATATCGGGAGTTTAAATTGCTCC-3 ' human ERBB2IP SEQ ID NO: 63 SEQ ID NO: 64 5'-GGGGAGAGCTACGTGGTAGTATG SEQ ID NO: 65 5'-CCAATACTTTCAAAGAAAACTT-3 ' human ELMO2 SEQ ID NO: 66 SEQ ID NO: 67 5'-CTCTAGAAGACCCCAGGAAGTTGGCCTT-3 ' SEQ ID NO: 68 5'-GAATTCCCCCAAGGTTCCATCCTAAGA-3 ' human BAP1 SEQ ID NO: 69 SEQ ID NO: 70 5'-CTCTAGACCTGACTCTGCAGCCCACTCTT-3 ' SEQ ID NO: 71 5'-GAATTCAGGAAGAGCTGAGTGGTGCCAG-3 ' human WDR37 SEQ ID NO: 72 SEQ ID NO: 73 5'-TCTAGAGTCTGGGTGTCTTAGGATGATGG-3 ' SEQ ID NO: 74 5'-TCTAGAGTTCAATTTCTAGCTCATCGTGATG-3 ' human KLHL20 SEQ ID NO: 75 SEQ ID NO: 76 5'-TCTAGAAGAAAAATCTTTGAATAAGACTAAC-3 ' SEQ ID NO: 77 5'-TCTAGAAGCTAAAATTTTTTTAATGGTAA-3 ' human PTPRD SEQ ID NO: 78 SEQ ID NO: 79 5'-CTCTAGACGTTATGTGGCTTTGCCATCC-3 ' SEQ ID NO: 80 5'-GAATTCGTAACAAATGGAACTTGTGTCAGC-3 ' human RASSF2 SEQ ID NO: 81 SEQ ID NO: 82 5'-TCTAGATGTGTGCACACATGGTACGTGTC-3 ' SEQ ID NO: 83 5'-TCTAGATGTATGGCTCCCTTGCTGAGG-3 ' human VAC14 SEQ ID NO: 84 SEQ ID NO: 85 5'-TCTAGAAACACTAAGGGTCGTCACGCC-3 ' SEQ ID NO: 86 5'-TCTAGACTTTGTGCTGGTGCCTACGTG-3 '

gene GenBank Access Number Cloning site of 3'UTR Vector designation Drosophila gene ush NM_057432.2 4154-4168 pAC- ush atro NM_206303.1 6524-8298 pAC- atro CG8445 NM_176194.1 1718-2091 pAC- CG8445 cbt NM_164384.1 1495-2003 pAC- cbt Lap1 NM_137163.3 3817-4029 pAC- Lap1 Ced -12 NM_135704.2 2285-2451 pAC- Ced- 12 CG12333 NM_142466.2 1761-1948 pAC- CG12333 dbo NM_080250.2 2416-2604 pAC- dbo Lar NM_078880.2 6218-6458 pAC- Lar Human genes FOG2 NM_012082.2 3494-4322 pAC- FOG2 SEPT7 NM_001788.4 1518-2410 pAC- SEPT7 RIN2 NM_018993.2 2738-3963 pAC- RIN2 SHC1 NM_003029.3 1552-2860 pAC- SHC1 HOXB5 NM_002147.3 877-1759 pAC- HOXB5 KLF11 NM_003597.4 1704-3867 pAC- KLF11 TCF7L1 NM_031283.1 1851-2717 pAC- TCF7L1 ERBB2IP NM_018695.2 5022-6826 pAC- ERBB2IP ELMO2 NM_133171.3 3235-3554 pAC- ELMO2 BAP1 NM_004656.2 2317-3244 pAC- BAP1 WDR37 NM_014023.3 3627-4589 pAC- WDR37 KLHL20 NM_014458.3 3229-3462 pAC- KLHL20 PTPRD NM_002839.2 9771-10044 pAC- PTPRD RASSF2 NM_014737.2 1436-5354 pAC- RASSF2 VAC14 NM_018052.3 2649-3040 pAC- VAC14

<8-2> miR -8 and / or miR -200 miRNA Regulation of target genes

Each reporter plasmid and Renilla luciferase expression vector of Table 4 (Promega, USA) and miR-141, miR-200a (hsa-miR-200a, SEQ ID NO: 87: 5'-UAACACUGUCUGGUAACGAUGU-3 '), miR-200b, miR-200c (has-miR-200c, SEQ ID NO: 88: 5'-UAAUACUGCCGGGUAAUGAUGGA-3 ') and miR-200 family of miR-429 and miR-8 (dme-miR-8, SEQ ID NO: 89: 5'-UAAUACUGUCAGGUAAAGAUGUC Each miRNA of -3 ') was purchased from Samchully and transformed together with Drosophila S2 cell line or human Hela cell line, and then luciferase activity was measured using a dual-luciferase assay ( Promega, USA). ) And the results are shown in Table 5. Seven gene pairs were selected that responded to both miR-8 and mir-200 family miRNAs in Drosophila and human cells, respectively (FIG. 7B).

※ Ts: Targetscan; pt: Pictar; mr: Miranda; mt: miTarget; mc: microT

※ Average of multiples of expression levels in at least two independent experiments

※ Values reduced by 0.8 times or less are indicated in bold.

※ n.d .: Not detected

Example 9 . Ush , the target gene of miR- 8 , regulates body growth

<9-1> miR -8 Identifying Phenotypes Represented by Target Genes

The following experiment was performed to confirm whether the target gene selected in Example 8-2 has a physiologically corresponding activity with the phenotype observed in miR-8 null fruit flies. Specifically, mir8 prepared in Example 6-2 The target gene in Drosophila CgG4 ush, Lap1, Ced -12, CG8445, and CG12333 and, the UAS vector (Vienna RNAi Library Centre) expressing the dsRNA of the (known that prevent neurodegeneration), the contrast gene atro mir8 CgG4 larvae were transformed in the same manner as in Example 6-2 to ush , Lap1 , Ced- 12, CG8445 , CG12333 And atro are fat-specific knockdowns of mir8 Cg > ush i, mir8 Cg > Lap1 i, mir8 Cg > Ced-12 i, mir8 Cg > CG8445 i, mir8 Cg > CG12333 i and mir8 Cg > atro i transformed Drosophila were prepared.

It was confirmed by quantitative-RT-PCR whether each gene actually knocked down in the knockdown fruit flies. Specifically, by extracting total RNA from 03 larvae (96 hours after incubation has passed) of the transgenic fruit flies using a pair of primers for each gene shown in Table 6 in the same manner as in Example 4 ush, Lap1, Ced - 12, CG8445 , CG12333 Or quantitative PCR for atro mRNA was performed respectively. At this time, the rp49 gene was PCR together as a quantitative control using the primer pair for rp49 of Example 4. ush , Lap1 , Ced- 12, CG8445 , CG12333 and atro mRNA expression level is After converting the relative value according to the amount of rp49 mRNA, Ct (comparative cycle threshold) value was analyzed according to the manual (User Bulletin 2, Applied Biosystems, USA). Repeating the experiment three times, and then, are shown graphically in the average value (FIG. 8b). As a result of the quantitative-RT-PCR, it was confirmed that other genes except Lap1 were knocked down (FIG. 8B).

gene primer Sense primer Antisense primer ush SEQ ID NO: 90 5'-ATTAAATCGGAGCCTCTGGA-3 ' SEQ ID NO: 91 5'-GGCTGTCGCTGGCCT-3 ' Ced -12 SEQ ID NO: 92 5'-GTGCCGGAAAACTCATTC-3 ' SEQ ID NO: 93 5'-GTACAGCTGGTGGGTCATCT-3 ' CG8445 SEQ ID NO: 94 5'-CATGAATCACGGCGG-3 ' SEQ ID NO: 95 5'-TGAACTTATCGTAGTTGTGGGT-3 ' CG12333 SEQ ID NO: 96 5'-GGACTTCCGAGAGGCAAT-3 ' SEQ ID NO: 97 5'-ATCGCAGTCCAGCAGC-3 ' Lap1 SEQ ID NO: 98 5'-CACAACAAGTGCAATATACGG-3 ' SEQ ID NO: 99 5'-TGCTATTCAAAGCATCTTGGT-3 ' atro SEQ ID NO: 100 5'-AGCAGGATACGCCCAAC-3 ' SEQ ID NO: 101 5'-TGCGGCTACCGGACT-3 '

<9-2> miR -8 miRNA Is the target gene for ush

CgG4 in Table 1 (wild-type), a mir8 prepared in Examples 6-2 CgG4 (miR-8 board Drosophila) and mir8 Cg> mir8 (that miR-8 null a fat body-specific expression of the miR-8 in the fruit fly Drosophila melanogaster) and, a mir8 prepared in Examples 9-1 Cg > ush i, mir8 Cg > Lap1 i, mir8 Cg> Ced-12 i, mir8 Cg > CG8445 i, mir8 Cg > CG12333 i and mir8 The weight of Cg > atro i Drosophila strains was measured in the same manner as in Example 3, and when the miR-8 null Drosophila expressed miR-8 in a fat-specific manner, it gained weight similar to that of the wild type. The same effect was seen in the case of knocking down ush specifically for fat body in 8 null fruit fly (FIG. 8A). This confirmed that the target gene of miR-8 miRNA was ush.

It has been reported that miR-8 targets atrophin ( atro ) to prevent neuronal degeneration (Karres et al . , 2007). The present inventors performed the following experiments to determine whether atro is expressed in fat body and is involved in body regulation. atro is known that according to the atlas fruit fly (環椎, Atlas, the first cervical vertebra), expressed in fat body in larvae (Chintapalli, VR et al ., 2007, Nat Genet 39, 715-720). But fat atro Knockdown did not heal the small body phenotype of miR-8 null fruit flies (FIG. 8A). From this, the function of miR-8 in preventing neurodegeneration (Karres et. al . , 2007) are separated from the molecular level as well as the functions and spaces for body growth.

<9-3> ush  Phenotyping of Knockdown Drosophila

We ush by dsRNA Ush1513 Drosophila (Cubadda, Y et. al ., 1997, Genes Dev 11, 3083-3095) was crossed with wild type fruit flies (w ¹¹¹⁸ ) or miR-8 null fruit flies (mir-8 ^Δ2 ) to ush1513 Heterozygote (w ¹¹¹⁸ ush1518) and miR-8 board ush1513 A release conjugate (mir-8 ^Δ2 ush1518) was prepared. Mir -8 ^Δ2 and ush1513, respectively Flies were crossed with double balancer fruit flies to produce fruit flies with each mutation in one individual using standard breeding methods. W ¹¹¹⁸ , w ¹¹¹⁸ ush1518, mir-8 ^Δ2 and mir-8 ^Δ2 ush1518 Drosophila body size was measured and compared in the same manner as in Example 3, ush1513 Heterozygote had a larger adult population than wild-type Drosophila, and a small population phenotype was relieved when ush was expressed at low levels in miR-8 null Drosophila (FIG. 8C). It was found that USH inhibited body growth.

Example 10 . Confirmation of the inhibition of USH expression by miR- 8

<10-1> miR -8 board In Drosophila ush mRNA Expression level

The present inventors confirmed by performing quantitative RT-PCR whether the expression of mRNA of USH selected as a target gene of miR-8 is actually increased in miR-8 null Drosophila. Intrinsic ush by performing quantitative RT-PCR in the same manner as in Example 4 above on whole RNA isolated from larvae or fat bodies of larvae mRNA levels were determined. At this time, primer pairs of an ush sense primer (SEQ ID NO: 102: 5'-CGAATTCCGATGTATCCAACG-3 ') and a ush antisense primer (SEQ ID NO: 103: 5'-GGAGTTGTTGTTCTCGTGCACC-3') were used, and the rp49 of Example 4 was used. Using primer pairs, rp49 was PCR together as a quantitative control.

As a result, ush mRNA was more expressed in the larval fat body (~ 2.0 times) than in the whole miR-8 null larvae (~ 1.3 times), indicating that miR-8 inhibits ush in fat body (FIG. 9A). This shows that ush is more strongly inhibited in fat than other body parts.

<10-2> miR -8 board In Drosophila USH  Confirmation of Protein Expression

The inventors performed a Western blot to see if the expression of the protein of USH selected as the target gene of miR-8 is actually increased in miR-8 null Drosophila.

In addition, Western blot was performed in the same manner as in Example 4 using an anti-USH antibody (Peptron, Korea) in the larva or fat body of the larva. At this time, by using the anti-ACTIN antibody to observe the expression level of ACTIN was used as a quantitative control.

As a result, USH protein levels were observed to be significantly reduced than mRNA levels (FIG. 9B), indicating that miR-8 inhibits USH expression through both destabilization and translation inhibition of USH mRNA.

<10-3> miR -8 ush  3 ' UTR of miR  Confirmation of control over the target site

<10-3-1> mutation ush  3 ' UTR miR  Vector Production with Target Sites

The present inventors performed the following cloning to determine whether a change in regulation by miR-8 occurs when mutations are introduced into the miR-8 target site of the 3 'UTR of ush .

Specifically, the PCR products for the 3 'UTR region of the ush obtained by PCR in 8-1, respectively, were digested with Bam H1 and Sac I restriction enzymes, and then pGL3_CMV vector (modified from Promega's pGL3) was digested with the same restriction enzymes. , Kim et al . , Nucleic Acid Research, 2009) were prepared by inserting a reporter plasmid having a miR-8 target sequence in the 3 'UTR sequence of the luciferase gene in, "Luc- ush 3'UTR ” .

In addition, the Luc- ush Using the primer pair and the QuickChange Site-Directed Mutagenesis kit (Stratagene , USA), SEQ ID NO: 104 in the 3'UTR (5'-CAACCATCGACGACAACTCTCCCGGAAAATCTCC-3 ' ) and SEQ ID NO: 105 (5'-GGAGATTTTCCGGGAGAGTTGTCGTCGATGG TTG-3 ') of ush 3, by introducing a point mutation in the target site of miR-8 UTR "Luc- ush 3'UTR mut ”.

In addition, the genomic regions of miR-8 miRNA obtained by PCR in Example 6-1 were respectively cut by Eco RI and Xba I restriction enzymes, and then inserted into pAC 5.1 vectors (Invitrogen) cut by the same restriction enzymes. The “Ac mir8 ” vector was prepared.

<10-3-2> ush  3 ' UTR Mutation miR  Introduced at the target site MiR in mutants -8 Check whether it is adjusted

In Drosophila S2 cells pGL3_CMV vector (Luc null, positive control), Luc- one prepared in Example 10-3-1 ush 3'UTR or Luc- ush 3'UTR muts were co-transformed with Ac mir8 and renilla luciferase expression vectors (Rehwinkel J et al. , RNA, 2005) using the known DDAB method (Han, 1996), and then luciferase activity was observed. Measurements were performed on a luminometer (Biorad) using a dual-luciferase assay (Promega, USA). As a result, it was confirmed that when the miR-8 target site included in the 3 'UTR of luciferase was mutated, luciferase activity was increased by the positive control (FIG. 9C). This indicates that miR-8 binds directly to the predicted target site of and mediates expression inhibition. The predicted target site for miR-8 is D. virilis, which is lineage distant . And Drosophilid species, including D. grimshawi . This shows that ush is the true target of the miR-8.

Example 11 . Confirmation of PI3K Regulation by miR- 8 and USH

Several studies have shown that larval fat insulin signaling regulates organism growth in an involuntary way (Britton et. al . , 2002; Colombani et al . , 2005). In order to confirm this, the inventors conducted the following experiment to confirm whether the growth of fruit flies is inhibited when insulin signaling is inhibited in the larval fat body. Specifically, the CgC4 Drosophila of Table 1 was crossed with either a PI3K DN Drosophila (Drosophila expressing a dominant negative PI3K) or a dsInR Drosophila (Drosophila expressing a dsRNA for InR) to Cg> PI3K DN (fatty-specific dominant negative PI3K Expression) and Cg> dsInR (fatty expression specifically expressing dsRNA for InR). CgC4, Cg> PI3K DN and Cg> dsInR Drosophila body size was measured and compared in the same manner as in Example 3, and as a result, Cg> PI3K DN and Cg> dsInR. Drosophila had a smaller adult population than CgC4 fruit flies. In addition, CgC4 and Cg> PI3K DN The cell size of the fruit fly wings was measured and compared in the same manner as in Example 3, but there was no significant difference, and CgC4 and Cg> dsInR The number of wing cells of Cg> dsInR Drosophila was smaller than that of CgC4 Drosophila when the cell numbers of the wings of the fruit flies were measured and compared in the same manner as in Example 3 (FIG. 8C).

Example 12 . Confirmation of Specific Disruption of Insulin Signaling in the Fat Body of miR- 8 Null Drosophila

When we inhibited insulin signaling in Drosophila fat, we found that Drosophila appeared similarly to miR-8 null Drosophila (FIG. 1D), with a smaller body size (FIG. 10A) and fewer flies (FIG. 10B). Assuming that it would inhibit body growth by inhibiting insulin signaling, we examined whether insulin signaling is defective in the fat bodies of miR-8 null Drosophila. Specifically, tGPH vector (vector expressing GFP protein fused to PH domain, Britton et al . , 2002) were transduced and expressed in wild type larvae or miR-8 null larvae in the same manner as in Example 6-2, and then dissected the third larvae of each group in the same manner as in Example 5 in fat body. Immune staining was performed. In this case, Zamboni's fixative (4% paraformaldehyde, 7.5% saturated picric acid in PBS) was used for tGFP staining, and anti-GFP antibody (Sigma, USA) and anti-FOXO antibody (1: 200, from O. Puig) Obtained). Fatty tissue samples of the wild type larvae or miR-8 null larvae were stained with phalloidin and DAPI to observe actin and nuclei, respectively. Since the PH domain binds to membrane-attached PIP3 (membraneanchored phosphatidylinositol-3,4,5-triphosphate) produced by PI3K, the PI3K activity can be measured by detecting membrane-attached PIP3 via the GFP signal expressed by tGFP. have. We found that PI3K activity was down regulated in the fat body of miR-8 null Drosophila as compared to wild type (FIG. 11A) to further increase the nuclear localization of FOXO (FIG. 11A). In addition, anti-p-Akt (phosphorylated-Akt) antibody (Ser505, Cell Signaling), anti-Akt antibody (Cell Signaling, US) in the fat tissue of wild type larva or miR-8 null larva ), Western blot using anti-p-JNK (phosphorylated-JNK) antibody (Santa cruz, USA) and anti-JNK antibody (Santa cruz, USA). Compared, there was no difference in the level of phosphorylated-JNK, but phosphorylated-Akt was significantly reduced (FIG. 11B). This suggests that insulin signaling is specifically disrupted in the fat body of miR-8 null larvae.

Example 13 . miR -8 activates PI3K to promote cell growth in a cell-voluntary way

<13-1> miR By -8 PI3K  Confirm activation

In order to further understand the function of miR-8 in adipocytes, the present inventors prepared a mosaic clone overexpressing GAL4 when a miR-8 is expressed in a fat body of a miR-8 heterozygous conjugate using a known method (Seth S. Blair, Generating Imaginal Disc Clones in Drosophila, CSH Protocols , 2007; pdb.prot4793). Anti-GFP antibody (cell membrane staining, PI3K activity measurement) and anti-CD2 antibody (miR-8 null cell staining) after transformation of tGPH vector in the same manner as in Example 12 above in mosaic adipocytes overexpressing miR-8 As a result of immunostaining, the GFP signal was increased in the cell membrane of miR-8 overexpressing cells, and the cell size was larger than that of miR-8 null cells (FIG. 12). This suggests that miR-8 promotes cell growth by activating PI3K in a cell-voluntary manner.

<13-2> miR -8 Fat body  Confirmation of Tissue-specific Cell-voluntary Growth Promotion

The inventors next tried to observe the loss of function phenotype through a twin-spot analysis (mitotic clone analysis). Mitotic null clones of miR-8 were prepared using the FLP / FRT system (Golic, KG . And Lindquist, S, 1989). At this time, when the thermal shock is given, chromosome recombination occurs specifically, and after cell division, one cell becomes a null clone and the other cell becomes a twin-spot having two phenotypes. Cells of miR-8 null clones were smaller than the surrounding cells of twin spots (cells obtained from miR-8 wild type copy) (FIGS. 13A and 13B). This indicates that miR-8 also promotes fat cell growth, as predicted that miR-8 enhanced insulin signaling in fat bodies. The inventors found that when clones were induced in embryonic or newly hatched larvae, few null clone cells were generated near the twin spot cells. This means that failure of proliferation / survival of miR-8 null cells is frequent during larval development (FIG. 13A). When twin-spot analysis was performed on wing or eye discs in the same manner as above, miR-8 null clones were produced, but growth inhibition of the organs was minimal (Karres et al. al . , 2007) (FIG. 13C). This suggests that the effect of miR-8 on cell growth depends on tissue morphology, which may be explained by the fact that USH is present in fat body but not in wing precursor cells or eye discs (Cubadda et. al . , 1997) (FIG. 13D).

Example 14 . Identify the role of ush in insulin signaling

<14-1> Insulin Signaling USH Check the role of

The present inventors prepared a mosaic clone of adipocytes overexpressing USH in the same manner as in Example 13-1 to confirm whether USH negatively regulates insulin signaling. In the USH-overexpressing mosaic clone, the tGPH vector was transformed in the same manner as in Example 12, followed by immunostaining using anti-GFP antibody (cell membrane staining, PI3K activity measurement) and anti-LacZ antibody (USH expressing cell staining). As a result, USH-overexpressing cells were smaller in size compared to the surrounding wild-type cells, showed a significantly lower tGPH signal in the cell membrane (FIGS. 14A and 14B), and a high FOXO signal in the nucleus (FIG. 14C).

<14-2> In the insulin signaling pathway USH The adjustment position of the

The present inventors prepared mosaic adipocytes expressing dsRNA for ush in the same manner as in Example 13-1 to observe the ush knockdown phenotype. In the mosaic ush mutant cells, tGPH vectors were transformed in the same manner as in Example 12, followed by immunostaining using anti-GFP antibody (membrane membrane staining, PI3K activity measurement) and anti-CD2 antibody (miR-null cell staining). As a result, it was confirmed that the tGPH signal was significantly increased (FIG. 14D) and the FOXO signal of the nucleus was decreased (FIG. 14E). From this, it was found that USH inhibits insulin signaling in parallel with or above PI3K in cellular spontaneous methods.

Example 15 . Confirmation of Inhibition of Insulin Signaling of Fatty Body of miR- 8 Null Larva by Increased ush Expression

Excessive insulin signaling is known to reduce the levels of insulin receptor ( Inr ) and cytohesin Steppke ( step ) through negative feedback of FOXO (Fuss et. al . , 2006; Puig and Tjian, 2005). We used CgG4 , mir8 to confirm that reduced insulin signaling when miR-8 is not expressed is recovered through knockdown of USH. CgG4 , mir8 CgG4 > mir8 And after the total RNA was isolated from the fat cells of mir8 Cg> ush i Drosophila and quantitative RT-PCR for Inr and step mRNA, the target genes of FOXO was performed in the same manner as in Example 4. The primers used at this time are shown in Table 7. As a result, the two targets of the FOXO were upregulated in the fat body of miR-8 null larvae, it was confirmed that the expression is rapidly reduced by the re-introduction of miR-8 (Fig. 15). Especially, mir8 Cg > ush mRNA levels of Inr and step , FOXO target genes, in the fat body of i (Fig. 15). From this, it can be seen that the increase in ush expression in the fat body of miR-8 null larvae at least partially affects the deficiency of insulin signaling.

gene primer Sense primer Antisense primer Inr SEQ ID NO: 106 5'-AACAGTGGCGGATTCGGTT-3 ' SEQ ID NO: 107 5'-TACTCGGAGCATTGGAGGCAT-3 ' step SEQ ID NO: 108 5'-CACCAGGAACTTCTTGAATC-3 ' SEQ ID NO: 109 5'-TTCTTTGTACCAGGATGTCA-3 '

Example 16 . FOG2 , a target gene of miR- 200 , regulates PI3K in human cell lines

<16-1> fog2  3 ' UTR Mutation miR  Introduced at the target site MiR in mutants Check the adjustment by -200

Ush miRNA target gene search results of Example 7 ush mRNA had a predicted binding site of one miR-8, while ush 's mammalian ortholog , FOG2, had a predicted binding site of at least three miR-200 family miRNAs (FIGS. 16A and 16B). We believe that Drosophila miR-8 miRNA is ush Just as the direct binding to 3 'UTR of mRNA regulated expression, the following experiment was performed to confirm whether human miR-200 family miRNA also directly regulates 3' UTR of fog2 mRNA to regulate expression. Specifically, in order to obtain the cDNA of FOG2 described in SEQ ID NO: 110, the total RNA was extracted from the K562 cell line using RNeasy mini kit (QIAGEN, USA), and then 1 μg total RNA and 1 μl Oligo-dT (Invitrogen, After filling up to 50 μl of distilled water, the mixture was placed in a SuperScript First-Strand Synthesis System (Invitrogen, USA) for RT-PCR reaction. CDNA was synthesized by reacting at 65 ° C. for 6 minutes, 4 ° C. for 5 minutes, 42 ° C. for 60 minutes, 95 ° C. for 5 minutes, and 4 ° C. for 5 minutes. Using the cDNA (3-5 μg) as a template, a primer pair of SEQ ID NO: 111 (5'-GGATCCATGTCCCGGCGAAAGCAAAGC-3 ') and SEQ ID NO: 112 (5'-GCGGCCGCTCATTTGACATGTTCTGCTGCATG-3') and a PCR Mastermix (Promega, USA) PCR was performed. After completion of the reaction, the PCR products of FOG2 generated were digested with restriction enzymes of Bam HI and Not I, respectively, and then pCK-flag vector (Heo I et. al ., Cell, 2009) to prepare a "pCK-FOG2" vector. One (FOG2 m1, FOG2 m2, FOG2 m3), two (FOG2 m12) to three miR-200 target sites of the FOG2 3 'UTR using the pCK-FOG2, the primers of Table 8 and the QuickChange Site-Directed Mutagenesis kit , FOG2 m23) or three (FOG2 m123) mutations were introduced into the Hela cell line with the expression of wild type FOG2 (FOG2 wt), FOG2 m1, FOG2 m2, FOG2 m3, FOG2 m12, FOG2 m23 or FOG2 m123. After transfection with the vector and the synthesized siGFP, miR-141 / 200a or miR-200b / c / 429, luciferase activity was measured in the same manner as in Example 8-2. The same amount of miRNA containing the sequence was mixed and used as a result, fog2 When all the estimated target sites of the 3 'UTR of the mRNA were mutated (FIG. 17, m123), the expression of luciferase was not inhibited by the miR-200 family miRNA. Thus fog2 It suggests that the inhibition of miR-200-mediated FOG2 expression is regulated through the predicted target site present in the 3 'UTR of mRNA. On the other hand, miR-200a and miR-141, which have one nucleotide mismatch in the seed sequence, inhibited luciferase expression, although less effective than other members. This suggests that non-regular target sites can also cause decreased expression of target genes (Bartel, DP 2009, Cell 136, 215-233; Brennecke, J et. al ., 2005, PLoS Biol 3, e85).

<16-2> Confirmation of Correlation Between Expression of FOG2 Protein and miR-200c Clustered miRNA by Tissue

FOG2 is expressed in the heart, brain, testes, liver, lungs and skeletal muscle (Holmes et al . , 1999; Lu et al . , 1999; Svensson et al . , 1999; Tevosian et al . , 1999). Despite relatively widespread expression in adult tissues, little is known about the function of FOS2 compared to its role in embryonic heart development (Fossett and Schulz, 2001). On the other hand, miR-200 miRNA is known to be expressed in various adult organs, including the pituitary gland, thyroid gland, islet, testis, prostate, ovary, breast and liver (Landgraf et al. al . , 2007). We performed a correlation between expression of FOG2 protein and miR-200c cluster miRNA in human cell lines from different organs, respectively, by performing Western blot and Northern blot for expression of FOG2 protein and miR-200 family miRNA, respectively. Confirmed. Specifically, anti-FOG2 antibody (Santa cruz) and anti-GAPDH antibody (Santa cruz) in HepG2, Hep3B, Huh7, FAO, AsPC1, PANC1, SW480, HCT116, HCT116p53KD, MCF7, MDAMB231, U2OS and Hela cell lines Western blot was performed in the same manner as in Example 4. In addition, Northern blot was isolated in the same manner as in Example 5 after separating total RNA from HepG2, Hep3B, Huh7, FAO, AsPC1, PANC1, SW480, HCT116, HCT116p53KD, MCF7, MDAMB231, U2OS and Hela cell lines using Trizol reagent. Was performed. At this time, oligonucleotides corresponding to miR-141 and miR-200c were end-labeled with [γ ³² -ATP] and used as probes of the northern blot. As a result, miR-200c cluster members and expression levels of FOG2 generally had a negative correlation supporting the inhibitory role of miR-200 in FOG2 regulation (FIG. 18).

gene primer Sense primer Antisense primer m1 SEQ ID NO: 113 5'-TTAATTTATTTTACCAGTGACATTCATAGCTGTGGTT-3 ' SEQ ID NO: 114 5'-AACCACAGCTATGAATGTCACTGGTAAAATAAATTAA-3 ' m2 SEQ ID NO: 115 5'-ACAAAATTACTGGAAGTGACATTGTAAGTAATGAGG-3 ' SEQ ID NO: 116 5'-CCTCATTACTTACAATGTCACTTCCAGTAATTTTGT-3 ' m3 SEQ ID NO: 117 5'-TTGTAAGTAATGAGGTGACGTTAATCAGTTTTATCT-3 ' SEQ ID NO: 118 5'-AGATAAAACTGATTAACGTCACCTCATTACTTACAA-3 '

Example 17 . Confirmation of specific regulation of PI3K - Akt - FOXO signaling by miR -200

<17-1> FOG2 Association between insulin signaling in humans

We have found that miR-200 miRNA actually decreases FOG2 and that Drosophila human miR-200 has a conserved role in the regulation of insulin signaling as in the case of miR-8 miRNA, miR-200 and FOG2 Is transformed into 30 nM of synthesized siGFP, miR-141 / 200a or miR-200b / c / 429 in Huh7 cells (FIG. 18), a hepatocellular carcinoma expressing relatively low but detectable levels, followed by anti-FOG2 Western blot was carried out in the same manner as in Example 4 using the antibody, anti-p-Akt antibody, anti-Akt antibody and anti-GAPDH antibody. As a result, FOG2 protein levels were significantly reduced (FIG. 19A). In addition, 2'-O-methyl oligonucleotide antisense (2'-) against siGFP, miR-141 / 200a or miR-200b / c / 429 was applied to AsPC1 cells, a pancreatic cancer cell line expressing relatively high levels of miR-200 miRNA. Treatment with O-methyl oligonuclelotides antisense (Table 9, Samchully) at 200 nM was performed by Western blot in the same manner as described above. As a result, FOG2 protein levels were increased (FIGS. 19B and 18). The results suggest that FOG2 is an aggressive target of the intrinsic miR-200 miRNA. When the miR-200 family was transformed, the phosphorylation of Akt was increased as compared with the transduction of the negative control group (FIG. 19A), and when the miR-200 family inhibitor was transformed, Akt phosphorylation was decreased (FIG. 19B). ). In addition, after treatment with siGFP or siFOG2 (SEQ ID NO: 119: 5'-UUAUUCAAGUCCAUCUUCCdTdT-3 ', Samchully) to the FAO cell line was confirmed by Western blot in the same manner as described above, FOG2 knockdown increased p-Akt levels , miR-200 family miRNA showed similar effects (FIG. 19C).

Sequence number Base sequence anti-siGFP SEQ ID NO: 120 5'-AACAUCGCCAUCUAAUUCA-3 ' anti-miR-141 SEQ ID NO: 121 5'-CCAUCUUUACCAGACAGUGU UA-3 ' anti-miR-200a SEQ ID NO: 122 5'-ACAUCGUUACCAGACAGUGU UA-3 ' anti-miR-200b SEQ ID NO: 123 5'-UCAUCAUUACCAGGCAGUAU UA-3 ' anti-miR-200c SEQ ID NO: 124 5'-UCCAUCAUUACCCGGCAGUAUUA-3 ' anti-miR-429 SEQ ID NO: 125 5'-ACGGUUUUACCAGACAGUAUUA-3 '

<17-2> FOG2 The role of insulin in signaling

We transformed the pCK-flag (empty vector) and the pCK-FOG2 prepared in Example 16-1 to the Hep3B cell line, and then analyzed the FOG2 for PI3K activity by immunocomplex kinase analysis using an antibody against p85. The effect of (Fig. 20) was confirmed. Specifically, pCK-flag (empty vector) and pCK-FOG2 prepared in Example 16-1 were transformed into Hep3B cell line, followed by a medium without serum or a medium containing 100 ng / ml of IGF-1. I replaced it. After 24 hours, proteins were isolated by treatment with lysis buffer (137 mM NaCl, 20 mM Tris-HCl, pH 7.4), 1 mM CaCl2, 1 mM MgCl2, 0.1 mM sodium orthovanadate, 1% NP-40). The samples were treated with anti-p85 monoclonal antibody (Santa Cruz) to immunoprecipitate PI3K. Precipitated antibody-PI3K immune complexes were mixed with protein A-Sepharose bead (Pharmacia, Sweden), followed by two 1, wash buffer solutions (0.1 M Tris-HCl ( pH 7.4) with lysis buffer solution. ), 5 mM LiCl, and 0.1 mM sodium orthovanadate). PI3K activity was determined by 60 μl of the protein-A Sepharose beads / antibody-PI3K immune complex, 10 μg of sonic-crushed PIP2 (Calbiochem, USA) and 1 μl of [γ- ³² P] ATP (500 μCi / ml). The kinase assay buffer (containing 10 mM Tris-HCl ( pH 7.4), 150 mM NaCl, 1 mM sodium orthovanadate and 10 μl of 100 mM MgCl 2) was analyzed. After reaction at 37 ° C. for 20 minutes, the reaction was terminated by adding 20 μl of 6N HCl. Lipids were extracted with CHCl3: MeOH ( 1: 1) and analyzed using a scintillation counter. in vitro For PI3K activity analysis, the immune complexes were incubated with purified His-labeled FOG2 protein at 4 ° C. for 2 hours.

As a result, when FOG2-expressing plasmids were transduced in Hep3B cells, insulin like growth factor-1 (IGF-1) failed to induce PI3K activity, suggesting that POG2 inhibits PI3K (FIG. 20). .

In addition, in the cell line treated or not treated with IGF-1 after transformation under the same conditions as described above, Western-based anti-p-Akt antibody, anti-Akt antibody and anti-GAPDH antibody were used. As a result of the blot, when FOG2 was treated in IGF-1-treated cells, phosphorylation of Akt was reduced compared to the case where no FOG2 was treated (FIG. 21A).

<17-3> miR The effects of -8 on insulin signaling pathways

We analyzed the effect of miR-200 miRNA on the downstream transducer of Akt (FIG. 21). Since Akt inhibits FOXO activity, we found the luciferase reporter plasmid (pFK1tk-luc) with 8 FOXO binding sites (Biggs, WH et al ., 1999, Proc Natl Acad Sci USA 96, 7421-7426) was used to determine the level of FOXO activity in cultured cells. Specifically, siGFP, miR-141, miR-200a, miR-200b, miR-200c, miR-429 or siFOG2 synthesized in Hep3B cells were transformed with pFK1tk-luc vector, followed by Example 8-2. Luciferase activity was measured in the same manner. In addition, an oligonucleotide having a sequence complementary to luciferase, miR-141, miR-200a, miR-200b, miR-200c, or miR-429 is transformed into a Hep3B cell with a pFK1tk-luc vector, followed by the same method as described above. Luciferase activity was measured by. In addition, pCK-flag (empty vector) and pCK-FOG2 prepared in Example 16-1 were transformed into Hep3B cell line, and luciferase activity was measured by the same method as described above. Luciferase activity in Hep3B cells was significantly reduced by transduction of miR-200 miRNA and FOG2 knockdown (FIG. 21B). In addition, FOXO activity was increased in the cells by treatment of miR-200 inhibitors (FIG. 21C). Correspondingly, FOXO activity was increased when FOG2 was overexpressed (FIG. 21D).

<17-4> miR By -8 FOG2  Identify the effect of expression regulation on insulin signaling pathways

The present inventors transformed pCK-flag (co-vector) or pCK-FOG2 prepared in Example 16-1 to HepG2 cell line to determine whether miR-200 specifically affects the insulin signaling pathway. After replacement with a medium without serum or a medium containing 100 ng / ml of IGF-1, anti-p-ERK antibody (Sigma, USA), anti-ERK antibody ( Sigma, USA) and Western blots performed with anti-GAPDH antibodies showed that, unlike the components of the PI3K signaling pathway, phosphorylation levels of Erk were similar with or without treatment with FOG2 (FIG. 21A). In addition, after transforming siGFP, miR-141 / 200a or miR-200b / c / 429 synthesized in the Huh7 cell line, nothing was processed, or Ly294002 (PI3K inhibitor), SB600125 (JNK inhibitor) or U0126 (Mek1). Inhibitors) were treated with 10 μM each, and FOXO activity was measured in the same manner as in Example 17-3. As a result, inhibitors of JNK or Mek1 / 2 did not affect miR-200-mediated FOXO regulation, but did not reduce FOXO activity by miR-200 when treated with PI3K inhibitors (FIG. 21B). From the results, it was found that miR-200 specifically regulates PI3K-Akt-FOXO signaling.

Example 18 . Function as voice adjuster of PI3K FOG2

Since stimulation of PI3K and AKT promotes cell proliferation and inhibits apoptosis (Pollak, 2008), we found that MTT (3- (4,5-dimethylthiazol-2-yl) -2,5 in Hep3B cells. Cell viability was measured by -diphenyltetrazolium bromide analysis. Specifically, after dispensing Hep3B cells into a 96-well plate, transformed with 30 nM of synthesized siGFP, miR-141, miR-200a, miR-200b, miR-200c, miR-429 or siFOG2, and then 50 μl of MTT solution (2 mg / ml) (Promega, USA) was dispensed into each well and incubated at 37 ° C. for 1 hour. Then, 150 μl of DMSO was added to each well, and then measured using a microplate reader (Bio-rad, USA) at a wavelength of 590 nm. In addition, 200 nM oligonucleotide having a sequence complementary to 200 nM luciferase, miR-141, miR-200a, miR-200b, miR-200c or miR-429 was transformed into Hep3B cells, and then the same as above. MTT analysis was performed by the method. As a result, the introduction of miR-200 miRNA increased cell viability (FIG. 22A) and the introduction of the inhibitor of the miRNA showed the opposite effect (FIG. 22B).

To identify the mechanism of action of FOG2, we performed immunoprecipitation against p85 in Hep3B cell line followed by Western blot for p85, p110 and IRS-1 (FIG. 23). In particular, when FOG2 was expressed, the amount of p110 and IRS-1 precipitated with p85 decreased. From this, it was found that FOG2 acts as a negative regulator of PI3K by interfering with the formation of the IRS-1 / p85p110 complex.

Example 19 . FOG2 directly binds to p85α to inhibit PI3K

<19-1> in cells FOG2  And PI3K Confirm interaction of

Although FOG2 is thought to be a nuclear transcription co-regulator, some studies have reported that FOG2 is also located in the cytoplasm (Bielinska et. al . , 2005; Clugston et al . , 2008). In order to confirm the above results, the inventors separated the nuclear and cytoplasmic fractions from Hela or PANC1 cells, and then separated the proteins from each fraction to obtain anti-FOG2 antibodies, anti-Lamin antibodies (nuclear fractions) (Santa Cruz, USA). ) And anti-Tubulin antibody (cytoplasmic fraction) (Santa Cruz), Western blot was performed in the same manner as in Example 4, it was observed in the cytoplasm more than the nucleus (Fig. 24a). DAPI staining in HepG2 cells and immunostaining in the same manner as in Example 5 using anti-FOG2 antibody and anti-p85 antibody showed that FOG2 is located in the cytoplasm (FIG. 24B), which shows FOG2 Suggests that it functions in the cytoplasm.

Given that FOG2 inhibits PI3K and colocates with p85 (FIGS. 17-25), the inventors wondered whether FOG2 interacted with PI3K. The present inventors performed immunoprecipitation in the same manner as in 17-2 when PANC1 cells were treated with or without 1 μg (Santa cruz), and then the anti-FOG2 antibody and anti-p85 antibody were used. Western blot was performed in the same manner as in Example 4 to confirm the expression of endogenous FOG2, and a significant amount of p85, a regulatory subunit of PI3K, was precipitated with the anti-FOG2 antibody (FIG. 27A).

<19-2> In a test tube FOG2  And PI3K Confirm interaction of

We transformed the Hep3B cell line with pCK-flag (empty vector) or pCK-FOG2 prepared in Example 16-1, and then with serum-free medium or a medium containing 100 ng / ml of IGF-1. After replacement, immunoprecipitation was performed in the same manner as in Example 17-2 using the anti-85 antibody, and then in Example 4 using the anti-p85 antibody, the anti-p110 antibody, and the anti-FOG2 antibody. As a result of Western blot in the same manner, the interaction of FOG2 and p85α was observed even when the expression of the two proteins was induced in FLAG-labeled form (FIGS. 26 and 25B).

Example 20 . Functions of the miR -8 / miR -200 and USH / FOG2 preserve the species

<20-1> p85 Combined with FOG2 Of domain Mapping

In order to map the domain of FOG2 that binds to p85, the inventors used the cDNA of FOG2 of Example 16-1 as a template, and each FPG using each primer for the truncated mutation of FOG described below. Amplified polynucleotides for cleavage mutations:

FOG2 [1-412] (SEQ ID NO: 126): sense primer (SEQ ID NO: 127: 5'-GGATCCATGTCCCGGCGAAAGCAAAGC-3 '), antisense primer (SEQ ID NO: 128: 5'-GCGGCCGCGTGGCTGGCTGTAAGCTGTC-3');

FOG2 [413-789] (SEQ ID NO: 129): sense primer (SEQ ID NO: 130: 5'-GGATCCCAGACTTATTGACCAGAAG-3 '), antisense primer (SEQ ID NO: 131: 5'-GCGGCCGCGATATCACATCTTGGGTGGTAG-3');

FOG2 [802-1151] (SEQ ID NO: 132): sense primer (SEQ ID NO: 133: 5'-GGATCCCTCTGACGATCAACAAGTG-3 '), antisense primer (SEQ ID NO: 134: 5'-GCGGCCGCTCATTTGACATGTTCTGCTGCATG-3');

FOG2 [1-506] (SEQ ID NO: 135): sense primer (SEQ ID NO: 136: 5'-GGATCCATGTCCCGGCGAAAGCAAAGC-3 '), antisense primer (SEQ ID NO: 137: 5'-GCGGCCGCTCATTTGACATGTTCTGCTGCATG-3'); And,

FOG2 [1-789] (SEQ ID NO: 138): sense primer (SEQ ID NO: 139: 5'-GGATCCATGTCCCGGCGAAAGCAAAGC-3 '), antisense primer (SEQ ID NO: 140: 5'-GCGGCCGCTCATTTGACATGTTCTGCTGCATG-3').

The PCR products obtained above were digested with restriction enzymes of Bam HI and Not I, respectively, and then inserted into pCK-flag vectors digested with the same restriction enzymes to express "pCK-FOG2_1-412", "pCK-FOG2_413-789", Expression vectors for FOG mutations of "pCK-FOG2_802-1151", "pCK-FOG2_1-506" and "pCK-FOG2_1-789" were prepared.

 Each of the mutants including FLAG-labeled at the N-terminus obtained above was expressed in HepG2, and then immunoprecipitated in the same manner as in Example 17-2 using an anti-FLAG antibody, followed by anti-p85. The antibody and anti-FOG2 antibody were used for the same analysis as in Example 4 above. As a result, it was confirmed that the intermediate region of FOG2 (507-789 aa) mediated interaction with p85α (FIG. 26).

<20-2> Confirmation of Inhibition of PI3K Activity of p85 Binding- FOG2 Domain

When the mutant FOG2 proteins prepared above were expressed in HepG2 cells, the inventors confirmed whether the intermediate region was sufficient to inhibit PI3K (FIG. 27A). As a result, the N-terminal site or C-terminal site had no inhibitory effect in PI3K activity, but the intermediate region had a significant PI3K inhibitory effect (FIG. 27A).

<20-3> Direct binding of conserved FOG2 and p85 between species

In order to test whether FOG2 directly binds with p85, the present inventors digested the PCR product for the p85 gene obtained in Example 20-1 with BamHI and EcoRI, and then the pGEX5X-1 vector digested with the same restriction site. (GE Healthcare, USA) to prepare a GST-fusion recombinant p85 expression vector. After transforming the expression vector to BL21 (Promega, USA) by electroporation, the protein FOG2 protein was expressed and purified in bacteria using a known method (Studier, FW et al ., 1986, J. Mol . Biol . 189 , 113-130), using the purified GST-fusion recombinant p85 protein in Used for in vitro binding assays. The recombinant FOG2 protein comprises an intermediate region of FOG2 (413-789 aa) that specifically binds to recombinant p85 (FIG. 27B).

The inventors have found that FOG2 utilizes recombinant FOG2 in vitro As with PI3K analysis, it was confirmed that p85 was directly inhibited. The activity of PI3K was significantly reduced when recombinant FOG2 protein comprising the middle region (with tro PI3K assay) was added to the immunoprecipitated PI3K complex (FIG. 27C). These results suggest that FOG2 binds directly to p83, causing inhibition of PI3K activity. In particular, we found that Drosophila USH physically interacts with Drosophila p60 (dp60, Drosophila ortholog of p85α) when dp60 is co-expressed in USH and human HEK293T cells (FIGS. 28A and 28B). This suggests that the mechanism of action of USH / FOG2 is preserved beyond species (Figure 29).

1 is a miR-8 null (null) of fruit flies (mir -8 ^{△ 2)} and weight of the female and male phenotypes in wild-type adult flies (a), (b), pupation and emergence (c), and wings cell size ( It is a figure comparing d).

Figure 2 is a Western blot confirming the activity and gene expression of the protein involved in insulin signaling in miR-8 null Drosophila (a) and the result of confirming the expression level of 4EBP via Q-PCR in miR-8 null larva (b) ).

Figure 3 miR- via immunostaining in fat bodies, cuticles and brain tissues of stage 3 larvae (96 hours after spawning) of mir -8 enhancer trap GAL4 strain Drosophila ( mir -8 gal4 ). Figure 8 shows the results of confirming the spatial expression pattern of (a) and the result of confirming the miR-8 expression amount through Northern blot in the whole larva, fat body, intestine, brain and cuticle (b).

4 mir8 CgG4 > mir8 and mir8 pplG4 And weight comparison of wild-type fruit flies (a) and (b) mir8 elavG4 and mir8 The figure which shows the result of comparing the weight of elavG4 > mir8 .

5 is a diagram comparing the homology of human miR-200 family miRNA and Drosophila miR-8.

6 is a diagram showing the results of confirming that the miR-200 family promotes cell proliferation in human cell line MCF7 (a) and a schematic diagram of an algorithm applied to the selection of target genes and orthologs using a target prediction program (b) .

Figure 7 shows the target gene candidate (a) for Drosophila miR-8 and human miR-200 miRNA selected using miRanda, miTarget, microT, PicTar and Target Scan, and the 3'UTR site of the gene candidate was luciferase. (B) shows the results of selecting seven gene pairs that are applied to the expression system and respond to both miR-8 and mir-200 family miRNAs.

8 is mir8 Cg > ush i, mir8 Cg > Lap1 i, mir8 Cg > Ced-12 i, mir8 Cg > CG8445 i, mir8 Cg > CG12333 i and mir8 As a result of measuring the weight of Cg > atro i transgenic Drosophila (a), it was confirmed by quantitative RT-PCR whether each gene knocked down in the Drosophila (b) and w ¹¹¹⁸ , w ¹¹¹⁸ ush1518, mir-8 ^Δ2 and mir-8 ^Δ2 ush1518 The figure showing the result of measuring the body size of the fruit fly (c).

9 shows that the expression of mRNA of USH selected as the target gene of miR-8 in miR-8 null Drosophila is actually increased by quantitative RT-PCR (a), and Western blot whether the expression of USH protein is actually increased Results (b) and the regulation of the miR-8 ush 3 'UTR miR target site was confirmed through the introduction of mutations in the miR-8 target site by measuring luciferase activity (c) to be.

10 is a diagram showing the results of measuring the body size (a) and the wing cell number (b) of the fruit fly when insulin signaling was inhibited in the fruit fly fat body.

FIG. 10 shows the results of observing PI3K activity and FOXO nuclear localization in miR-8 null Drosophila fats to determine whether there is a defect in insulin signaling in the fat body of miR-8 Null Drosophila (a) and wild-type larvae or miR Jb and Akt phosphorylation level in the fat tissue of the -8 null larva was confirmed by Western blot (b).

12 is a diagram showing the results of observing PI3K activity and cell size through immunostaining in mosaic adipocytes overexpressing miR-8.

FIG. 13 shows that twin spots of miR-8 null clones are generated more slowly than wild type clones (arrows) and have a lower DNA content through Twin-spot analysis (mitotic clone analysis) (a). (3) As a result of confirming PI3K activity in adipocytes and wild-type cells overexpressing USH (b) When the miR-8 null clone was generated by twin-spot analysis in the wing or the eye disc, the result confirmed the growth inhibition of the organ (c) and (d) show the results of confirming USH expression in wing progenitor cells and eye discs in Drosophila larvae.

14 shows the results of confirming cell size (a), PI3K activity (b) and FOXO signal (c) in the nucleus by immunostaining in USH overexpressing cells, and PI3K activity (d) and nuclei in USH knockdown mosaic adipocytes. Shows the result of confirming the FOXO signal (e).

15 shows CgG4 , mir8 CgG4 , mir8 CgG4 > mir8 And mir8 Cg > ush i Inhibition of insulin signaling of the fat body of miR-8 null larva by increased fat cell ush expression in Drosophila was performed by performing quantitative RT-PCR on Inr and step mRNA, target genes of FOXO . to be.

16 is ush As a result of searching for a predicted binding site of miR-8 in mRNA (a), a figure showing a result of searching for a predicted binding site of miR-200 family miRNA in FOG2 (b).

Figure 17 shows the introduction of mutations in one (FOG2 m1, FOG2 m2, FOG2 m3), two ((FOG2 m12, FOG2 m23) or three (FOG2 m123) to three miR-200 target sites of the FOG2 3 'UTR. When, it is a diagram showing the results confirmed by measuring the luciferase activity whether regulation for miR-200 miRNA.

Figure 18 shows the correlation between the expression of FOG2 protein and miR-200c cluster miRNA in human cell lines from different organs, respectively, by Western blot and Northern blot for expression of FOG2 protein and miR-200 family miRNA in each cell line, respectively. The figure which showed the confirmed result.

19 is a result of confirming the expression level of FOG and the degree of phosphorylation of Akt when transforming miR-141 / 200a or miR-200b / c / 429 in Huh7 cells, which are hepatocellular carcinoma (a), and the pancreatic cancer cell line AsPC1 When cells were treated with 2'-O-methyl oligonuclelotides antisense against miR-141 / 200a or miR-200b / c / 429, the expression level of FOG and the degree of Akt phosphorylation were measured. As a result of Western blot (b), and when the FAO cell line treated siFOG2, the expression of FOG and the degree of phosphorylation of Akt was confirmed by Western blot (c).

20 is a diagram showing the results of confirming the effect of FOG2 on PI3K activity by transforming pCK-flag and pCK-FOG2 in Hep3B cell line, and through immunocomplex kinase analysis.

Figure 21 transformed pCK-flag and pCK-FOG2 in the Hep3B cell line, and then confirmed the degree of Akt phosphorylation according to whether IGF-1 and FOG2 treatment by Western blot (a), eight FOXO binding sites As a result of measuring the activity change of luciferase according to the transduction of miR-200 miRNA in cells transfected with luciferase reporter plasmid (pFK1tk-luc), and (b) and transduction of complementary oligonucleotide As a result of measuring the change in activity of luciferase according to (c), and FOG2 overexpression in Hep3B cell line is a diagram showing the result of measuring the change in activity of luciferase.

FIG. 22 shows the introduction of miR-200 family miRNA (a) and complement to miR-200 family miRNA through MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) analysis in Hep3B cells Figure 4 shows the results of measuring cell viability according to the introduction of the oligonucleotide (b).

Figure 23 shows the results confirmed by immunoprecipitation analysis of the formation of IRS-1 / p85 p110 complex when FOG2 is expressed or not expressed in Hep3B cell line.

Figure 24 shows the results of Western blot confirming the expression of FOG2 in the nuclear and cytoplasmic fraction of Hela or PANC1 cells (a) and the results of confirming the position of FOG2 by immunostaining in HepG2 cells (b).

25 is a diagram showing the results confirmed by immunoprecipitation analysis whether FOG2 and p85 are located together in PANC1 cells.

FIG. 26 shows the results obtained by immunoprecipitation analysis of FOG2 mutations that bind to p85α when each FOG2 mutation and p85α are expressed together in human cell lines derived from various tissues.

Figure 27 shows the results of measuring the PI3K activity according to the increase in the amount of FOG2 expression treated with mutant FOG2_1-412, mutant FOG2_413-789 and mutant FOG2_802-1151 in PANC1 cells (a), GST-fusion recombinant p85 protein and mutant FOG2_413- 789 or mutant FOG2_802-1151 directly binds in Results (b) confirmed through in vitro binding analysis, and when FOG2_413-789 or mutant FOG2_802-1151 is added to the immunoprecipitated PI3K complex, it is a diagram showing the result of confirming the PI3K activity (c).

Figure 28 shows the results confirmed by immunoprecipitation analysis that the physical interaction of fruit flies USH Drosophila p60 (dp60, Drosophila ortholog of p85α) when dp60 is expressed in USH and human HEK293T cells together Figures: (a) IP with USH, (b) IP with dp60.

29 is a schematic diagram showing the intracellular signal transduction process of USH / FOG2.

<110> SNU R & D Foundation <120> Screening Methods of Agents for Regulating the miRNAs and their          targets which control Insulin Signaling Pathway <130> 9p-09-47 <160> 140 <170> KopatentIn 1.71 <210> 1 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> siGFP <400> 1 ugaauuagau ggcgauguu 19 <210> 2 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-141 <400> 2 uaacacuguc ugguaaagau gg 22 <210> 3 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-200b <400> 3 uaauacugcc ugguaaugau ga 22 <210> 4 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> has-miR-429 <400> 4 uaauacuguc ugguaaaacc gu 22 <210> 5 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> 4ebp sense primer <400> 5 atgcagcaac tgccaaatc 19 <210> 6 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> 4ebp antisense primer <400> 6 ccgagagaac aaacaaggtg g 21 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> rp49 sense primer <400> 7 agggtatcga caacagagtg 20 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> rp49 antisense primer <400> 8 caccaggaac ttcttgaatc 20 <210> 9 <211> 23 <212> DNA <213> miR-8 miRNA genomic sequence <400> 9 taatactgtc aggtaaagat gtc 23 <210> 10 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> miR-8 miRNA sense primer <400> 10 tcacaaagaa ttccttatcg cca 23 <210> 11 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> miR-8 miRNA antisense primer <400> 11 gctctagaat tttcagccgc ttgtcttcgc 30 <210> 12 <211> 931 <212> DNA <213> miR-200c cluster <400> 12 cggcgggccg acggtcgagg ggcttcggag ggcctgcttg gactgcaacc tgggcctcgt 60 gatcagcgac ccagggtgtg gctggtggcg ggcagcaggg ctcaccagga agtgtcccca 120 gggactcggg tggtgggggg atgggagcca gggatctgca gcttttccgc agggatcctg 180 ggcctgaagc tgcctgaccc aaggtgggcg ggctgggcgg gggccctcgt cttacccagc 240 agtgtttggg tgcggttggg agtctctaat actgccgggt aatgatggag gcccctgtcc 300 ctgtgtcagc aacatccatc gcctcaggtc cccagccctt agctggctgc agccccctcc 360 ccacttccca cgcaccccgg aagcccctcg tcttgagctg agagcgttgc acaaggggtg 420 gttcttgttg gctggctgcc actaagggac acaatgggcc ccagcccctc ctcccaccca 480 gtgcgatttg tcacctggtg gatccagaac ccacagtcga ccttgagctt ggggttggct 540 cgccccctct caagagacct cacctggcct gtggccaggg tcccctgtag caactggtga 600 gcgcgcaccg tagttctctg tcggccggcc ctgggtccat cttccagtac agtgttggat 660 ggtctaattg tgaagctcct aacactgtct ggtaaagatg gctcccgggt gggttctctc 720 ggcagtaacc ttcagggagc cctgaagacc atggaggact actgaccaac aacctctgac 780 cttcacccct ctggatgggg gacgaatcac taggcaaagg ggaacaatgg gaaggagaca 840 aaatggctgc ctttacagct gcagcaagat gtggaaacac tggttcccta ggcacctcca 900 ttgtcttccc gccttgggag catgaaataa a 931 <210> 13 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> miR-200c cluster sense primer <400> 13 cggaattcac ggtcgagggg cttcggag 28 <210> 14 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> miR-200c cluster antisense primer <400> 14 gctctagaca tgctcccaag gcgggaagac 30 <210> 15 <211> 4732 <212> RNA <213> Drosophila melanogaster ush mRNA <400> 15 caguuggcag cggaccgagc gacgagcgag agaguuucuc ggcugcugau acgaauucca 60 guccgagucc cgguaccagu ucgaauccca aagauaaaca acuuuuggcc ggcgucccgg 120 caucaacaaa caucagcugu gcugcgucgu caucgcaucg ccgcucgguu ugguucgcuc 180 gguaucugua ucgguuguau cuguaucuca cagcggcgca cgaaaauuuc auuacucacc 240 gccgcacuca gccggucggu ccgagucuca guuugcugug aaacgcggag cgagcagcaa 300 ccacgagcgg uacgcgguac uaaaaaucgu cagcggccga gggauuaaga gcuacaauac 360 gauaccgcac guucgucaua acgagaauuu gaauaauugu auuacaguga guagugaguc 420 aaagaaaauu cccacgggaa gugacaacau aauugccgcu uaaguacggc ccaaaccgaa 480 agauagaaau gugcaaguga caaagugaag gagccaccag uaaaccugga uuauuugcca 540 aguggauuac aauacacaua uagggaaaac aaugcuuagc agcaauacaa gaggugauug 600 uuccgauacc gcagaagaga ugaccgucga cagcagagau uccaaagauc ugagugcuca 660 ggauauaggc gagcagaagc agcagcaaau ggaggaccag uuggaggauc aguugaacga 720 uuccagagac ccccaaaaca acaauaacaa uaucgaugac gacgcugaug aagaugcuga 780 guucgaggag ccagaaaagg ccaaucccca acaggaucag gauuugggag agacagagau 840 ggaacaggaa cacgaucuuc agcaggagga ucuucagcag gaacugccug caaacagccc 900 aagcacuccg cccaggaguc ccagcucucc ccagcugauu cccaagcugg aacagcccgc 960 uacgccgccc ucggagccag aagccucucc gugccccuca cccucaccau gucccacgcc 1020 caaguacccc aaagugcguc ugaaugcgcu ccuggccuca gauccugccc uuaagccaga 1080 ugccaaggag cugacccugc cggacucucg ucuccuggcu ccuccgccgc ucgucaagcc 1140 ggacacgcaa gcgcaaccgg agguagcaga accucugcua aaaccagcca gauuuaugug 1200 ccugcccugu gggauugcau ucaguucgcc cucaacucug gaggcccauc aagccuacua 1260 uugcucucau agaauuaagg acacggacga ggcuggcagu gauaagucag gggcaggagg 1320 aucgggagcc acagcuggcg augcagcagg guugaccgga ggcuccaccg agccaccagc 1380 caaaauggcc aggacaggaa agcaauaugg cugcacccag ugcuccuaca gcgcggauaa 1440 gaagguaucc cuaaaucgcc acaugcgcau gcaccagacu ucgccagcag cucccacacu 1500 ggccggcuug ccaagucuuc uccagaacgg aauagcgccg ccgggaguua cgcccaaucc 1560 gauggaggau agcucuaguc aacaaaccga ucgcuacugc agccacugug auauccgguu 1620 caacaacauc aagaccuauc gagcgcacaa acaacacuac ugcaguucgc ggcgaccaga 1680 gggacagcuc acgcccaagc cagacgccuc uccgggagca gguuccggac cggguuccgc 1740 aggcggaucc auaggaguau cagcccaggc ggcuacgccc ggcaagcuga guccgcaggc 1800 aaggaacaag acuccgacuc ccgcaauggu cgccgucgca gcugcagcag cggcugccgc 1860 ugccucccuc caggccacgc cgcauucgca uccaccuuuc cuggcacugc ccacccaccc 1920 gaucauaauu gugcccugcu cgcugauccg ugcagcuagc uuuauuccag gcccacugcc 1980 cacgccgaac ucgggaaucg ugaauccuga aaccaccugu uuuaccgugg acaacggaac 2040 gauuaaacca cuggccacug cccuaguugg cgccaccuua gaacccgaac gcccuucagc 2100 uccaucaucu gcugcugagg ccacagaagc caagaguagu ccuccggagc ccaagcgaaa 2160 agaggcaggg uuaacuagag aaucugcgcc ucucgaucuc ucgcugcguc gaucgccaau 2220 cacccugaau ucacugagcu ugcgucagcg ccaacugcgg aaugcucucu uagaugugga 2280 agaaguacuc cuugccggcg uaggaaccgg caaggagaac guugagacuc cgagaggagg 2340 uggaagugug acucccgagc aaaucguuug cgcucccucc cugccgagca guccuucgau 2400 gagucccucg cccaaaagac gagccaucag uccacgcagu uccggggcug guagcgccuc 2460 cuccauguca ccaccugguc ucaauguggc uguuccccau cugcucgaca ugcgcuccau 2520 gcuaccugcg gacuuuggac ucuccgaguc gcuguuggcc aaaaccaacc cggaacuggc 2580 ucugaaacug gcggcggcag cggccgcagc ugcuguggcg ggcaguagug gagcagcagc 2640 cuucccgccc gcuucucuuc cugcccaaac uaguaguggg aauccgggua guggaggauc 2700 ggcagguggc gcccagcagc cucagaucua ugugaaaaag ggcgugucca agugcaugga 2760 gugcaauauu guguuuugca aguaugagaa cuacuuggca cacaaacagc auuauugcuc 2820 ggcaagaagc caagaaggug cgagugaagu ggaugucaag ucggccguuu caccauccau 2880 cgcuggagca ggaggacugg gagccggagc ugcagaggcu gcuucgucgg uggaaaccac 2940 accgguggca uaucaacagu ugaucugcgc cgccugcgga auuaaguaca cuucccugga 3000 caaccugcga gcccaucaga acuacuauug ccccaaggga ggagcaguug cugccccugc 3060 cgcgacgccc acggauccug gccaacuggg caugccaaag gagaagugcg gaaagugcaa 3120 gacauugcau gaaauugguc ugccuugucc accacccguu gcgaauccuc ucgcagcgcc 3180 gacagucaau ccgcaacccg ccaccaauag ucugaacaag uguccuguuu gcgggguagu 3240 gagucccaca gcagcacugg ccaagaaaca cauggagaug cacggaacag uaaaggcaua 3300 ccgcugcagc aucugccagu acaaggggaa cacucuucgc ggcaugcgca cccacauucg 3360 aacccauuuc gacaagaaga cuagcgacgu aaacgaggag cuauacauga ccugcaucuu 3420 ugaagaggau gcuagcgcgc ugagucagga acugguuacu ccaacagggg caucuaccac 3480 aacuggacac gauuccaugg aucaucccuc gcagauguuc aacugugauu acugcaacua 3540 ugucuccacc uacaagggca augugcugcg ccacaugaag cugaugcauc cccauguggc 3600 caucaacucu cccuccauuu cccccgacac cagagaucaa gaugugacuu ccaaccccac 3660 uaccaaccaa cacucgaauu ccgauguauc caacggugaa gcacccaguu uccacauuaa 3720 aucggagccu cuggauccac caccgacugu gaaucuggug cacgagaaca acaacucccc 3780 gaucgccacg ccacauauua aggcugagcc aaucgaaguc ggagcggaug cugcgcccgg 3840 cgggcugguu ccgccgauga cuucuccgcu gggaaacagc agcagcgugg cugccgcagc 3900 agcugccgcc gccgagguga ugaagaagua cugcucaacc ugugacauau ccuucaauua 3960 cgugaagacc uaccuggcuc acaagcaguu cuacugcaaa aacaagccca uucgccccga 4020 ggccagcgac agccccaguc cgaaucaccu gggcggaggc guugccgugg gccugggcau 4080 uggcggccug gucggcggac acggccagca gaagaacaag gaaaaccugc aggaggcggc 4140 cauuugagaa agccagcugc ggugggaggc gcagcgcaag cagcuugagu gguauuacac 4200 gugcuucuaa gcagcagccc aacccaaucg aaacuaaucg aauccuagcc uccuuuaaaa 4260 auguacacua uccaaaccgc aaccaaccau cgacgacagu auucccggaa aaucucccgu 4320 cgcugcccuc cacuuccaac uccauuuggg gacugacucc uucgacuuac cgaucgauag 4380 aaguagauau auacaucgua cucccguaca uauacaguac accuaaagga auaacacaua 4440 gauacuuagg uacauuaauu uaggcgcaua cccauuaagu gggaaccaug acauauaaug 4500 caugccaaag cuuaccacuu agacagaaau uaaauugagg gcgagaaacu acaaguuugc 4560 aauggucugc ggaguagaag gaggcgaagu auuccaaugu agauccaaua cuauaaugua 4620 aauuacucgc uguuccccau aaagcgagca auucuaacug uguaauuuaa augaguacau 4680 uauuuauuaa gugugugcga caauaaaagu aauuaauuua uucaauauaa aa 4732 <210> 16 <211> 33 <212> DNA <213> Artificial Sequence <220> 223 ush sense primer <400> 16 atcgatgagc tcgaaagcca gctgcggtgg gag 33 <210> 17 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> ush antisense primer <400> 17 acgcggatcc ggcagcactc aaatcgttcg ttg 33 <210> 18 <211> 8151 <212> RNA <213> Drosophila melanogaster atro mRNA <400> 18 cccccccccc gcaguuuuug aaagggugac cgacggaugg cuguguuugu gcaaauuaac 60 cgaaauauac auuuccggcu acucgcugcc uccucgcuga gcaacaacua caaaaacaac 120 aacaccaaua acaacagcag gaacaacaau aucaagaaca acaggagcaa agcaacaaga 180 accaccacga ccgccgccca cccccuuuga aaauccaucg acacagcacc acaucaccau 240 cgucgucauc aucaucauca uccucagcaa cuuggagucu ggaaucugga gacggagacc 300 guagaccgga guuacuuugc cgccugcugc uggagagagg agcaucuucg ugcgggacgc 360 cucgacgaua cacaaccccg acuacggauu cauguugaau uuucauaucg uagagggcau 420 aaaaaggcuu aacacuggac cggaagcgug uuuuuaaagg gugauggcgg ccuccacuca 480 aggagaaauu cgaguggguc ccggccacca gguaaacgau gucuaugcaa aacugcccga 540 uuauaaucca aucucaagcu uccccaucga caaggaaacc gaugaacgug aacuagagga 600 aucaagaugg aguccaggcg uuguggccga uggcgacuug uuaauguucu ugcgugcggc 660 ucgauccaug gcugcauuuc aaggaaugug ugaugguggu uuagaagacg guuguuuggc 720 ugcuagucgc gacgacacua caauaaacgc acucgacgug cuccacgauu cuggcuacga 780 uccaggcaaa gcucuacaag cgcucguaaa gugccccguu ucgaagggca ucgacaagaa 840 guggaccgag gacgaaacaa agaaauucau caagggucug cgucaguuug ggaagaacuu 900 cuuccgcauc cauaaggacu ugcugccgca caaggacacg ccggagcugg ucgaguucua 960 cuaucugugg aagaagacgc ccggcgcgaa caauaaucgg ccacacaggc gacgccgcca 1020 aagcgcccug cgacgcaacc gugucacgcg ggccaacaac agcaacagca acacuccucc 1080 gaagaaggag gacacuccag aaccacaaac ugcgacgacg gcgacggcgg cggcaaccgc 1140 ggcguccgag acggcgaguc gcuccucgcc cgcugucucc aaggaggaga acagcucgcu 1200 caccgaggac gacgccagcg agugcgacag ugauucgagu cugacccaca aaagggauga 1260 aucacccuca aggaugagga cgcguaacaa gcaacagaac aacaacagca gcaccagcag 1320 cgguaacaac acggccggca acgguggcgg uaacgccaca uccauaagca gcggaucaac 1380 cggcggcggu gccgcuggcg gcaauaguuc gucuaaggau caaucagcca acgccguggc 1440 uaauggcaag cgacccaaga ggggcuccga aacaccggac guuuccggcg gagccucggu 1500 cgauaguccc aagacaccga cgaaggcugu ggccgagagu ucagccaaua agcgcaaggg 1560 uggcaagcag gauacgccca acaagaagaa gcgaacggag caggagucca acgagccaag 1620 cgcccaugag gagaaugcca ucaaggagaa gcgcaagcga ccggacagcc cgguugagag 1680 uaugaacucg gauagcagac cggauucagu gcucgacgau ggggaaucga auaccacgga 1740 caccaccacc gccgagcagc aguccacaaa ggacagcaag gagacgguca gcugcaagga 1800 ggagcgcgaa auggucacca acgaucugga ggccaaggcc gaggaaaagg ccaucaaggc 1860 agaggcuuug gccgaagaca gcaaggauag cgccaucaag aacauggacg aggagacaaa 1920 cauccaggcg ccuagcagug cagacacuag uuugguggau gguccuaauc ccaaugcccu 1980 gcccaguccg guagccgcac caaucacaau gaagguacca acaauugcca ccguugaggc 2040 gcugaacgcg uccgucgacc gcaaggaggc caucgagaag auggagucgu gcgacagcga 2100 uccggagaug cuuaaaaaac uggcaaccau uaagcaggaa guaucuccgc agcagcaaca 2160 gcacaugcaa cagcaaucac agcagcagau gcagcagcaa cucgcuccag uuggcauacc 2220 gcaaccuccg ucuugcccgc caucggaauc agucuauauc aaaaaagagc ccauggagga 2280 cucgauggac gccaccugca aucagaacag caacgaaccg caggaccuga aggugaagau 2340 cgagauuaaa aacgaggaug cauugaagca uagugcugga ggucugccgc cuucaggucc 2400 gugugcaccg ccuucagcuc uacauccgcu uuccggagcu ccgguagaga gcggccagga 2460 gccacugcac cugcaacaca ugccucaugg gcaaguaacg acgcaaccgc ccccuggcua 2520 ccuaauugau ggccagcuaa aguauggacc aucgggacaa ggcgugccuc cacagccacc 2580 acaacugcac agcgaugcgg cuggaggagu cagcggagca ccaccuggag cgccaaccac 2640 gccccagaag uauccgcccg agauggagau gaaguucgcu ccucaggauc ucaaguaucc 2700 cccaccgccg ccucuagaug cacucaagua cagccaggag augcaagcug cggcggcggc 2760 agcggcugcu gcuggcaaau acgauaugaa guauaugaug gaacagcagg gcaaguacaa 2820 uguggaguug ucagcugccc aucagccgcc aagcaagcca ggcuaccagg acucgcuuaa 2880 gauacccgau auuaagcccg guuucggcca ccugccgcac aacgugggcu cuccgcugga 2940 cgccgcccau aaauacggac cgccuccgac gucgcaagag ucccagcaac agcaacccca 3000 gccgccggca caucagguac cgccgggagc aacuccacca ccugguaucg ccaugcccaa 3060 gccgcacuau caacacgaug ugcaaacacc accguuggga cggcccuucg agccgaccgg 3120 acuuaugcuc aaguauggcg auccauuggc agccaaauac ggcccgcccc aggaucucaa 3180 guacccgaug ccuccggucu cucaggcggg accagcggac guaaagcccu auggcggcga 3240 gaaucugauc aaguccucac cguacggacc gccgccggag aguccuaucg augccucugc 3300 gcgcucuaca ccuggucagg acagccaggg cagcaauagc aauucacagc cgcccucaau 3360 gcccccgcaa ccgcagcagu uccagucgcc gcaucccucg ccgcauaugc cuucgccagc 3420 aggagguggu cuaccaccgg gaaugcaucc gcaaaaucuc auccacggcc cgccaccagg 3480 ugcagcgggu ggcagugguc cccagccacc uccaccgccc acaucgcuac accagccaac 3540 gcccacgucu gcagguccac ccagucugca acauggacua cauccuggcc aucaacacuc 3600 acagcugucu guggcaucau cgauaccgcc gagcucgauu ggaauuccuc ccacgcucuc 3660 gacuauggcg cccucgcaca ugcacccgca ccuucaucca caugcgcauc ugcagggucu 3720 ccaucggccg cacgaucugc cgcccaguau gcauccacau gcacccaugc cgcugucguu 3780 gcagggacau ccgcagcacg gacauggauu gccgcccucg cacacuucuc agcaacagca 3840 gcagcagcaa caacaacaac agcccggcgg accagcuggu acggugcgaa cuccgucacc 3900 agcccagcag ccgccgagau ccaugcacga uccgcaaucg ucucgagagc cgcccacauc 3960 gcagccuucg accacuaugg caggaucgag ugguccgggu ggaccaccgc cgcaacaguc 4020 gccgcaugcg caucgcacau caccguugcc agggcucgcg gguagugguc cuccaccccc 4080 gggacucauc ggucauccga uggccauaca cccgcaccua gcccacuugc cgcccggaca 4140 ucccgcucac gcagcgcugg cccauccugg acaccaucug cugucgcacu cgauagcggg 4200 cuuggguccu ggugguggac ccaucgcguu gcuggccggu cccggcgguc uuggugguau 4260 uccagagucc gcucuaaguc gccgcacccc gcccucacau cugccacacu cgcaugccuc 4320 uucggcccca cugacggcuc acucgguggc caguaugacg ucuaccagua ugucgcugac 4380 caccagcacg gugccaucgu ccgccuuuag ccgcgccagu cccagcguac agaucucgag 4440 caguggagga ggaccuucag ggcccggaag cguuggaccu ggaggaaugc caaacucguc 4500 ggcagcagcg gcugcugcgg cagcugcuca ucgggcagcc ucaccggcau ccagcguaag 4560 cagccuaagu cggcagaguc cgcugcaucc ggugccgcag ucgccgcuca gccaucaucc 4620 cucguccucu gcguuauccg ccgcagcagc ugcuguggcg gagcgggauc gacaugcgcu 4680 gaugcgucag caaucgccac acaugacucc acccccggug uccaacgccu cuuuaauggc 4740 gaguccucug agcaagaugu acgcuccuca accgggucag agaggcuugg gaacaucacc 4800 gccaccgcau uugcggccag gagcaucgcc gccggucauu cgucacccgc agaugccucu 4860 accguugcca cugauugcgc cuggcggagg aaucccgcag auuggagugc auccggguca 4920 gucaccguau ccgcacccgc uucugcaucc cucgguauuc uacucgccgc aucaccaucc 4980 auuuaauucg ccauauggcu augcgcccua ugguccugga uucccggcau acaugaagcc 5040 gccaccacag ccgggacagc ucgauccggc agccgugaug gcggcccacc augccggauu 5100 gcaaggaccg ccgccccagc agaugcgcca ggacgagcag aaugcagcgg ccgccgcugc 5160 acaagcagcu gcugagaaac aacaccaagc ggcugcagca gcggcagccc agcagcacaa 5220 ggcgccgcaa caacaacagc cuggcggaau gccacccaac aaaccgccga cgccaaagac 5280 gccacagggu ccgggcggug ggaugccccc aggaaugggu ggaccgggaa caccgacggg 5340 acugccgcca ggugccuauc cuggcagcca uaugccggga uauccacaag ggccgccuca 5400 ugggucaccc uuugcgccac aagaugguca gccucacggc uugaagccca caucgcacau 5460 ggacgcccug cgagcgcaug cacacucagc caacucggcg gguaugggcg guggacacca 5520 uccgacggag ccauugccca uugauauuga accggaucca gagccagaga uucccagucc 5580 aacgcacaau auaccacgug gucccagucc cgaagcaaaa ccggacgaca ccgaaugcca 5640 ucgcucucag ucugccauau uugugcgcca caucgaucgu ggggauuaca auucgugcac 5700 gagaacagau uugaucuuca agccgguggc cgacucaaag uuggcccgca agcgugaaga 5760 acgcgaccgc aagcuggccg aaaaggagcg ugagcggcga cagcagcagc agcaacaaca 5820 acagcagcag caacaacagc aagcagcggc cgcgcaacag gcggcacagc aagccaagau 5880 gaaggcugag cugaagccac cguaugcgga uacgccggca cugcgucaac uguccgagua 5940 cgcucguccc cacgucgccu ucagggaacu ggaggagauu aaaaacgcac aagcugcugc 6000 ggcgagucaa ucccgacuag auccgcacug gauggaauac uaucgacgcg gcauccaccc 6060 cucgcaguuc ccgcuguaug cgaauccggc gauaucgcag auggagaggg agcgucuggg 6120 aauuccaccu ccgcaccaug ugggguugga cccgggcgag cacauggugc guaugccgca 6180 accaccggag gccgguuucc aacugccacc gaauguuggc caguauccgc ggccaaauau 6240 gcuuauaccu agggagccgc acucggaugu ccugcugcgc auguccuaug ccgaccaacu 6300 acaggccgcc gaguuucagc gacagucccu gcaugaucag uacuuuagac aacggcccag 6360 auaaguggac agacagcaau uguagagagc aagcaacuga acaaagacaa uacucgggcc 6420 gcggccguga ggaacugcuu ugugugauuu uuguguucaa gcguuuacau uuuguuuucc 6480 acacgcacaa ccacccaaca uacucacaaa acaaacauca aaacaucaac aucaucugca 6540 gagacgacgu cagcguaugg cucaagaauc cagaaaccag aaaaccaagc gacuuuaagu 6600 ugcgcaacgc cuugucacgu uuuuauguaa aaagaaucag cugcguuaau cgggaguggu 6660 ucgucgauug uuccggcugc auacagcaca cuucacacca gacacucacu gaccaaggca 6720 uccagaauca aaaagcagaa accaacuaca aaacuaaacu aaacaaaaca ggacacgaug 6780 gucgcaauau cuuaaguauc cucuucauag uguauagugu uaaauucuua aacguaagcg 6840 agauauggag cauguuaaau acgguuuaga cuuauuaagg caauggaaca aaaucgaacu 6900 uggcacacaa auuuuuaugg agguuuucaa acagcuuaua uauuauaugu auaugaaugu 6960 gcuacacuuc gacaaaaacu aucuaguaag cuucacaaau caauugauuu agacacgaug 7020 auaacuaagc aaaaccaaaa cacaaacuca ccagcacuug cugcaggagg caagguacau 7080 aaauuaguau agacaaacga uuaaggauag acgaaucgag cgcucauaua uauauucgag 7140 aguauuguac aaauggauuc ggagauaguu uucuuuuuac aacacaguuu ggcacucugc 7200 gcauuagaga uacgccuaau auuauuuaaa uguauuuacu uccagccaca auugaacuau 7260 auaagguuua uauaugcuug cugcuuucga cuuuuuagcu aaaauaaaau gguaaacgaa 7320 ucacgccaau gccuagauuu cgcuuuuugu auguaguccu uguuuguguu agccuagcca 7380 gcagcuuugc cuaccaacua acugcuacuc auuauuuauu auuuugcuau gacuagagua 7440 uuauaggggu ugggauauga aaguuuugua aaaagugcau aaaaaauaac gaaauaauua 7500 uauggguauu augugaauga uuugaugcug cucucaaacc caaaagcaca gcacggcuuu 7560 cgaugcuugu gacgugccag cauuugaagc agcugcaauu uugagugcuu aauuuauauu 7620 aguaauuuau acaauuauua uacgcuaaaa aaugcauuga auucacacaa uauuucaaaa 7680 gauuuaguau guaaggccaa caaaccgcua agguggaugg auucaauggu aaacaaaagu 7740 gugaauuaua cuaaacaaaa ugagaaucca caaaguaaaa auauuuuuaa auacgguuuu 7800 uuauuucaca uuuugaauau acacauugau auaaauauau uguagcuaua uacuaauuau 7860 gaaaguuuau cguaggaguc ucgcaagcau auucaauggc acacaaacga caaaaaacaa 7920 aaaaaauaau acaagauuug gcguaguuaa cauuuaagaa guguaaagua aagauccaua 7980 cauauuuaua uauauauaca aaaaaaaaca aaacaaaugc agcuauauua uugcagcuau 8040 auuacuaaau aaauuauaaa acaaaaagau cauacugaaa agauaaauua uuaaaaccug 8100 uaaaaacuaa guaaaacaaa auaaaauuag uuacauuagu uaccaaaucc c 8151 <210> 19 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> atro sense primer <400> 19 atcgatgagc tcgtggacag acagcaattg tagagagc 38 <210> 20 <211> 35 <212> DNA <213> Artificial Sequence <220> <223> atro antisense primer <400> 20 acgcggatcc gcagagaagg ttttctggga tttgg 35 <210> 21 <211> 2121 <212> RNA <213> Drosophila melanogaster CG8445 mRNA <400> 21 cuaaggcaga uaagaggaag cgcaaaaaac uauuuugguc caaaucggua auauuuuuau 60 auuuaaaaug aauuggggac agaagauaac cggcagccga cggcccuaac uucuggacgg 120 acaucgccag uauauggauu gcuccuaaaa cuagcaaacc aacguacuua ugcucuaugc 180 aacaauaaau gucauaugaa aacaaacuau ugaagacguu uuuaugagua uaaccgauuu 240 ucauguguau auccuucucu gugcagaaaa acgaaauuaa aguaacaucu cggauacuuu 300 cgcagcaauu guauaagcca uacaaauaug aacgcugcug gaggagguag uggugcccag 360 gcggccgcag uggcggcagg caacaacagu cucagccaca augcgcuacu uuccacggcc 420 ucaggagcca ccaccaugcc uauggcccaa cuggcagacg guugguugga gcuggagucc 480 gacccagguc ucuucacacu gcuucucaag gauuuuggau gucacgaugu acagguggag 540 gagguguacg accuacagaa acccaucgag aguccauaug gcuucauauu ucucuuccgc 600 uggaucgaag agcgacgcgc cagacgcaaa auuguggaga caacugcuga gauauucguc 660 aaggaugagg aggccauuuc cagcauuuuc uucgcccagc agguaguccc caauagcugu 720 gccacacacg cguugcuuuc ggugcuccug aacugcaacg agaacaaucu ucagcugggc 780 gacacccuga gucgacuaaa aacccacacc aaaggcauga guccggagaa caaggggcug 840 gcuauuggca acacuccgga gcuggccugu gcccauaacu cacacgcaau gccgcaggcu 900 cggcgccguc uagaaaggac gggagcgggc gucucaaguu gccgcuucac gggugaggcg 960 uuucacuucg uuagcuuugu gccuaucaau ggacaacugu uugagcugga uggcuugaaa 1020 ccguauccca ugaaucacgg cgguugggag gauagcgagg auuggacaga uaaguuccga 1080 cgugugaugg cugagcgacu gggaauagcg accggcgaac aggacauacg uuucaaucuc 1140 auggccguag ugccagacag gcggauugcc auaacacaua aacuuaagau gcugcgcaca 1200 aaccaggcaa ucguguccgg aacacugcag aaacuucuaa aggccgacga gcagggcgaa 1260 ucggggaacg gagacucaca acguccggau acacccacca cauuauugga gccgagcgcu 1320 uucaccgcac gugaccugca aucgcuucuc aagaaccugg acacagaaau agccaucaau 1380 gagcagcauc uggcugacga aaacgaucgg aggcacaugu uuaaggugga cgccagucga 1440 cguacccaca acuacgauaa guucauuugc acuuuccucu cuaugcuggc ccaccagggc 1500 guccugggcg agcugguuag ccagcauuug cuuccgucga agaaggucag uggccagggu 1560 gcugccaauc ggaucagcaa gcagagcacc accgccagug caggcggaag caccgcagca 1620 ggcacugcau ccacgcccaa gacucagcaa cagcaggcug cggccgcuaa gaauggcaag 1680 ucgccaagca aaacgccagg caggaggcga aagggacgca acaagugcag aaaacggaag 1740 uaggcgacca gacgaccagc auguacaugu uuuuaagaau acaguuugca uuuuuaagac 1800 cauucauucc aguauuguuu uucuacaauu uccccaccaa gaagcaccau uaacccauaa 1860 gaucguauaa guuuaguuuu aguaacacuc aaaugcgguu cuuuuaaaua cgaaaucaac 1920 uuggcagcuu cauuuuuauu ugaucgcaau ucgaucuuuu uaauaauaaa aaaccauuuu 1980 guucguauga aucguuuuuc augugguccu ugauagacga aggacccaaa uguguaugca 2040 uuacauauga auaauauaua uauuuucuga uauauaucua uaugauaguu guauaacaaa 2100 aacguaaucc uucaauaauu c 2121 <210> 22 <211> 34 <212> DNA <213> Artificial Sequence <220> <223> CG8445 sense primer <400> 22 atcgatgagc tcccagacga ccagcatgta catg 34 <210> 23 <211> 35 <212> DNA <213> Artificial Sequence <220> <223> CG8445 antisense primer <400> 23 acgcggatcc caataacgtt aggattcacc cgatt 35 <210> 24 <211> 2044 <212> RNA <213> Drosophila melanogaster cbt mRNA <400> 24 auucgucucu ugaauuccga acgcaacggu ucgccuucgc uccaaaacgg caagcggcac 60 uugaaagcgc cagugcaaag uuagcugaag ugcaaaaauc uacacaaacu gagauuaaac 120 caagugauua gccccggaac aaaacaaaca aacaaacaca gacucuucaa uaaaaaaaag 180 agagugcaac ugaaauuuga aagugcaaug gacauggaua ccuugcuacc uucaccaccc 240 gcaacgcccc cacuaaggga aaacaaguug gaaaucgucg ccaaggacga acaacaggug 300 aacgagaauc ugcuaaaggc aaagcucaag cugguggccc aaaagaguca gaaaaaugga 360 gggauuauua caccaaaucc aucggacacg gaagacgagg cucccgaaau agcgguuccc 420 aacaagaaac cccguuugga acaaccugcu augagcauga caccgccgcc ggaucaaaag 480 cuggacgaug aucagaaagc ggaaaggguu agcgucauca ugcgggucaa caguucuggc 540 gcugucucuu cuaguagcca agacgagaac ucaucuaguu ccaccuccug cuguaguucc 600 ucuuccaaca caaacacaag uacaaguuca guaccaccca cuguggagga cgacuaucca 660 gaggccaaug uguggcgcaa ucucaaguuc aaaaugaaca gaaagcgugc ugcagaagug 720 gcacuacccc caguacaaac acccgagaca ccaguugcga agcuuguaac gccaccugcu 780 ccagcggaau gcaucaagga ggaggaaaua aaaccuauuc ugacaccgau auauguuagc 840 ccaguagcuu cgucugccag ccaacuuauc cugcucagca caguggccgc ccaacagagc 900 cccacacccg uacccaaaac gccaacaaug uccgaggaga aacuaacaac cagaauuacg 960 gccgcccagg cggcggccac cagaagucgc aucuacgagu gcaguuuucc cgauugcggc 1020 aagaauuacu ucaaaagcag ucaucugaag gcccaccaga ggguucacac cggcgagcga 1080 cccuuuaucu gcaaguggga gaacugugac aaacgauucu cccguuccga ugaguugucc 1140 cgacacaaac ggacccacac cggugaaaag aaauuccagu gcagcgucug ccagaagaaa 1200 uucaugagaa gcgaucaccu gucgaagcac gucaaacggc acaacaagga caaggcgaac 1260 ggagugaauc gacaugucuc ccuggccaac aacaacacau cagcuucggu agcggcuucc 1320 cucugcgaug ccucgcucca uuugcgagcg auagcaccgg cgggcuccag cgcuaguucc 1380 ucgcccauca guuccgccag cuugcaaguc uacagcgccc aagaucugcu gaggcuacag 1440 cagcaggcca gcaguuucac cuucggcgga acccugcuuc aagugcagcg augagaagag 1500 caaugcgagc agcaucagaa ccaaucucag ccccacagua ucgccagcua aacagagaug 1560 aauucccacc aaauuucaaa acaaauuaac aucugaguca aauucgaaau gccuucucgc 1620 ucucauguag cugguguaga cuuaagcuua guuaggagcu agaucuuguc accguuuugu 1680 gcauauauca acuucaaauu caauacguaa uacccaaauc ggauagaugc cacuucuuuc 1740 caggagcaca cagcacacuu cuuaucuaag cgaucuucuu cagauuaaua uucugaauau 1800 cccuaguguu uguacauacu auaaagagac gcauucggcg uccaacaucu gugauauuua 1860 aguuuucuua acucuuuucu auuguaccau aguugucagu guaauucucu uaagcuaaaa 1920 cacacacaau uguauuuauu gcuaaaugug aaaaaaagaa aaugaaaaca aauagaauaa 1980 uggaucaucg cacgauucca aucucaacaa augaaaauau acauauuuuc caaagaaaac 2040 uaaa 2044 <210> 25 <211> 34 <212> DNA <213> Artificial Sequence <220> <223> cbt sense primer <400> 25 atcgatgagc tcgaagagca atgcgagcag catc 34 <210> 26 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> cbt antisense primer <400> 26 acgcggatcc gattggaatc gtgcgatgat cc 32 <210> 27 <211> 4168 <212> RNA <213> Drosophila melanogaster Lap1 mRNA <400> 27 cggcauuuac caacacuaac accugcaagc aacuugucau uuucaaagcg aaaaacauua 60 aauaaaacgc aauuggaugg caaaaauaaa uauuaccccc ucauguuuga cauacagcgu 120 uugaguaagc aaugaaauca ucuauuuggg gacaguaaau gcgagaaaga gcgaaaagag 180 aggcucaguc agucgccucu guguguaugc gagugugggu gagugugccu gugcguguua 240 guguuggugg gcagaacaau aacaagccca aaaaugccaa uggacaugga auguaaugga 300 gcaggaccaa caauaacguu ccaucggagg cgaugaauca cagcagccac aaacgugccu 360 uaauuaacga acgaaaagca auacacuuaa uccaaagaug ccgcugcuga gcaaaugcuu 420 ccccugcuuc aaauucaagc gcgaggaggu gaucgacaag cuggacuaca guaauacccc 480 gcuaacggau uucccagaag uuuggcagca cgagcgaacc cuggaggagc uuuaccugag 540 caccacaaga uugcaagcuc ugccaccgca auuguuuuau ugccaaggau uaagggugcu 600 ucacguaaac agcaacaauu uggagagcau uccacaggcc auuggcaguc ugcgacagcu 660 gcagcaucug gaucucaacc ggaaucuuau ugucaaugug cccgaggaaa ucaaguccug 720 uaagcacuua acccacuugg accugagcug caacaguuug caacgucuuc ccgaugccau 780 uaccucgcua auuucgcugc aggagcugcu gcuaaacgag accuaucuag aauuccuacc 840 agccaacuuu ggccgauugg uaaaucugag aauucuggag cugcgacuca acaaucucau 900 gacacuuccc aaauccaugg ugagacuaau caaccuacag aggcuggaca uuggcgguaa 960 ugaauuuacc gaguugcccg aaguugucgg cgaguugaag ucccuucgcg aacuauggau 1020 cgacuucaau caaauacguc guguuucggc caacauugga aagcuacgcg aucugcagca 1080 cuucgaggcc aauggcaacu ugcuggacac ucuucccagc gagcugagca acuggcgcaa 1140 uguggaggua cuaucuauau gcuccaacag ccuggaggcc uuucccuuca gcgugggcau 1200 gcugaagucc cuggugacau ucaagugcga gucaaacggu uugacagagu uacccgacag 1260 cauaagcuac uuggaacagc ucgaggaacu aguauuaagc cacaacaagc uuauacgccu 1320 accuagcaca auugggaugu ugcguagccu ucgcuuucug uucgccgaug auaaucaacu 1380 acgacaacua ccagaugagc ugugcagcug ucagcaguug agugugcuaa guguggcgaa 1440 caaucaguug uccgcucuac cacagaacau uggaaaccug agcaagauga agguacucaa 1500 uguggugaau aacuacauaa augcauugcc aguuucgaug uuaaaucugg uuaaucucac 1560 aucgaugugg cuaagcgaua accaaucgca gcccuuggug ccgcugcaau aucuagaugc 1620 aaguacaaag acccaguuga ccugcuucau gcugccccaa gucacauuca agaugaacag 1680 uauacaggcc caacagcagg cucaggagca guaugaguuc guuuacgcca accagcagca 1740 gccucacgcg agucccucga gaaggauuug cuucgccgag gaggccacca uucuuagcaa 1800 cgccaaagca cagccagcgc ccaauuaucc cagcuuugug gccgcuccgc cgacgccaac 1860 gcccgaucag auggcugggu cugugcgguu aaugcguuca cccacaccuu aucccaagga 1920 gcugcgacaa auguccaaau augugaggca ggcucaggcg gcgaccucau cggccaaugc 1980 cagcgaggua agggaggcac gcguaguaac caauggacaa auucacugug auagcaacaa 2040 ugccaaccag gauguugugg accaggccac aacaagugca auauacggca uugcgcccga 2100 aacgacacac aucuacggag ucuaucagca accgcagcag auggcgcacc cggugccaac 2160 gcaggaguac uacggccugc cacuggucaa cuacgaagca cacuaucagc agcuuuacgu 2220 cgaggccaac acgccgcuuc ccaccacgca uuuaaacggg gaucaggacu augaguugca 2280 accgcugcag caacaaccaa ugcagcaaca ggcgcugcca acaccccggu uggaaccacc 2340 accauaccac auagcucgag uuuacacaaa gaaaacgccc gaggaucuca accuuuacga 2400 guccaugagg cagcgaaagc agcaacaaca gcugcaagaa caaaccaucu accaagaugc 2460 uuugaauagc aauaguaacu uuaaaacgac agcgauuggc gcucaggacg ucgaagaguc 2520 agucgaucag uuggacuacc aaaauaauau uagcaauaau uuagagccaa auccggagga 2580 ggaagaccag gagcuggaug acacuauguc gcagcacucg cuuaauucca cggccacuaa 2640 caauacuucc aaagcgagcc auaagaaauc caccuggauc uuuggugucc acaaaaaucc 2700 gacagucaaa cagguuacuc uaaaauggga gaacucuaua ggcuuugaua uagccgagcu 2760 ucuuaaucag guuggcaucu uugugagcuc cauaacgccc aauacgaaug ccgcccgccu 2820 gcuuaaucug aacgacaagu ugcuggaaau cgacggcuac gaccugacca augccaaccu 2880 gagcgaugcc aaacgagugc ugcuaaacug cggcacgguc augaacauaa uguugucgag 2940 aaaaugaugg accaucuauu uaccgaccua agccgauucc caucccgaga ccgaagacuu 3000 uuugugcauu uuucuaacgc auuuuuccaa uaagcagcau cgccgaaauc ggauaacaaa 3060 uuacuuggac cgaucauagu guuaguucau aauacccuuc cguacuggcg gucaaaccau 3120 cagucauaga gcacaaucgc cagcuagagg aucccaugcu uuuuguauau uagauagacg 3180 aauccaaacu gccauuuucu uugcaccucc auacucgaua gaagaauauu accagaagac 3240 ggccaaauuu aguaguuaua gcccacaaca aguauucuca aaauuuaagu ugaaugaaga 3300 ugaauugaau uuaugauuug acuuuaaagc uguuuauaau ucgaauugag ccacuuauaa 3360 guuggauuua uuuaaucagu aucuaaucga uaucuuaugu aaauaguuaa guuaugcacu 3420 uaaggaagca auaauauguu uggccaauua uuaguuccag uacuuuauua uuacaaugcg 3480 uagauuacau ugaaguggca uuuguauugg aaguuugugc uaccauuauu cugaccgcca 3540 guggaaugua ucagcauaau acauauaaau agugcaauua gcgucgauua uagguauaua 3600 uauacgcacu cauuauauau acucgcgaua aucagacaaa uuaccuucaa cuagcgccua 3660 uuuuacgccc agcguuguuc aaauuuguuc ccaaaauuau auguauagau caauuacuug 3720 auuacaaaag auauuugcug uuuauguuag uuguuauggu gcagcaauac aaauucuaua 3780 uauauauaua uuuacauaua cguacgcaau cuauauuuau uguccaaucg aucgaagugu 3840 caaacauuau uuucaauggu gaacuguuag cacaaacuga auucucacgc uauuggcauu 3900 gugaucagua uuauuauuua ucugguuaaa cacgugaaau cuaaucgaaa gauaagcacu 3960 ucuauauuca uagccuuuau auauauauau agucauacau agagaugugc auuauugaag 4020 aacacacgcc cacacacaca cacaaacaua cugcauaaau acgaaucuag cuaacuuaag 4080 caagcgcaca cauuuugcca auucaauuug auuuaagagc guuaccacaa aucacugaaa 4140 uacaacauuc acuuugucau uucgccau 4168 <210> 28 <211> 31 <212> DNA <213> Artificial Sequence <220> <223> Lap1 sense primer <400> 28 gagctcttat tgtccaatcg atcgaagtgt c 31 <210> 29 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> Lap1 antisense primer <400> 29 ggatccgcgt gtgttcttca ataatgca 28 <210> 30 <211> 2581 <212> RNA <213> Drosophila melanogaster Ced-12 mRNA <400> 30 acuacuuguc aauuggucac acugcacuga auuaaaaaaa aaaagaauau aaguuuacgc 60 agaaaaacaa uaauaauugc aauuaucgcu aauugaaaga gugccaaaau gauaccaaaa 120 aagacgacgg uuaaggauuc acacauugug aaaauagccg uggagcgaga ggaccauaua 180 gcgcaguuga uaaaucugga ucaaaggcau ccgcuggcaa guaaaaucca ggaaauuugc 240 aaugguuggu ccaucagcga ucaccagaac uaugcacucc aguucuacga gcccacuaac 300 cgaaaguacg ugaccgaaaa gaaucgcaau gagaucaaga auggcuccgu acugcagcug 360 caguauucgc cauccaagac ugccugcgau gccauggagg ugcugcucaa uggcagucca 420 caggagaagg cgcugcgucu aaaggagcuc acuucccuaa gcaccgauca cacauuugcc 480 cuggaguuca ucaaggagaa gggucucgac acacucauua agaugauuga ggauggcggc 540 cagaccaacg aagacauucu gaaauacagc cuggccagcu uuguggagcu gauggagcac 600 ggcacggugu ccugggaggu gccggaaaac ucauucgugg cccgcaacau ugagaucgug 660 cgaaacuuuc agaaauaucc uacgaacugu ggugagagcg cccuguccaa cuuggagaac 720 auuguuaugu gcagcaauaa acaugugcuu guugccgagg acaucaagcu gcaggauauu 780 cuccgauugc uucaggaugu caauucgccg gugaugcgac aaaaugccau agcccugcug 840 aacgcucugu ucgucaaagc agacgagggc cgccgcagga ccauugcuca aacuauuagu 900 gccaagcagu uccgccuggc ccuuaucggc aauggacugg gcacagagau gacccaccag 960 cuguacgugc ugcaaacguu gacucucggc cuguuggaga agcgcaugcg uaugaagaug 1020 aacgcgcagg accaggacgc gcacgagaag aucaaggagc ugcggcgcau ugccuucgac 1080 gauaauacca acgcuuugaa ucaaaacgac gaucacaucc gucguggcgg uggcuccgga 1140 gcuggcaaug ucaacuucuc gcaguacuac aaaaagcugg gcuucaagug cgauauuaac 1200 ccugcccagg auuucauuga gacgccgccg gguauauuag cguuggauug caugguguau 1260 uuugcacgca acuacacgca acaguaugcg aagaucguca gggagaacuc cugccgugcg 1320 gaugagcacg agugucccuu cggacgcacc uccaucgaau uggucaaggu gcuaugugac 1380 auccuucgca ucggcgagcc accagccgag caauccggug acuuucagcc cauguucuuu 1440 acgcacgauu cgccguucga ggaauucuuu ugcauaugcg ucauuacgcu gaaucgcaca 1500 uggaaggaua ugcgugccac ugccgaggac uucagcacaa ccuucagcgu ggugcgggag 1560 cagauccagc gcacucuaaa gggcagaccc gagaaccugg acgauuuccg aaacaagauc 1620 gcuuuguuga ccuaucagca gaucacaaca cugcgccagc aggagcgcac uucgaaggag 1680 gagugugauu ccacagcuuc ggcgauuguu aagcucaaag agaagauauc accgcaaauu 1740 cuugagcuaa ucaagcagca acggcucucu uuucuggugg aagguacaca uuuugcaaaa 1800 uacuugcgag gcacucgaac aaaggacaag uucugguacg cucgucuuuc gccgaaucac 1860 aaggugauuc auuacggaga uugcgacgag aagacaauac ccaccaugga ggaguugccc 1920 aagaaacugc ccaucaguga gauuaagcaa uugcuagagg gcaaggaaug cccgcauaug 1980 aaggaaacuc gcauucggaa aucggcggua aaccuggccu uuuccaucac cuucgaaaac 2040 auggagcacu cuacguugga cuuuguggca cccgacgaaa gcauauucaa cuacuggacg 2100 gauggaauaa acgcgcugcu gggucaaccg auggucagca agcaaaagaa cgaggacuuc 2160 gacacucugc ugucgaugga gaucaaauug cgccugcugg acacggaagg uguggacauc 2220 agcaaggacc caccgccaau accggaggau cccgaaaacu augauuuuug cuuugagagc 2280 uaagcauaac gagcacaauu acuucacgaa uuauucgauu cauucuagac ucccccauca 2340 cccaagauac augccaaagu guacacacaa cauaaucgua uucuccgauc ugaucuucua 2400 cuaguauuac uugugcaauu ccccgaacac aaaaugaaca aacuggugac ggucacgucc 2460 aagugacuaa ccgcugccua gccauuuagu gaacuuuaca ccauuuauga uuauaauagg 2520 uauauaauau auagagauca uuaaauaaca cauuuuauac auaacaauuu uguacaaaaa 2580 c 2581 <210> 31 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> Ced-12 sense primer <400> 31 gagctccata acgagcacaa ttacttcacg 30 <210> 32 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> Ced-12 antisense primer <400> 32 ggatcccgtc accagtttgt tcattttg 28 <210> 33 <211> 2168 <212> RNA <213> Drosophila melanogaster CG12333 mRNA <400> 33 acaaaaacac cugcaauaag uauauuauca uugcgaaaac caagugcaac agcaguuccc 60 cagucagcca aagaggcaaa ugcaucgcca gcagcggaag auggaucaau agagcgcgag 120 ucugcaaccc cugaacacua auuggauucc cccgggcgcu cugcccacca ggaaacggau 180 aucgccgacc agcucggcuc agaaaaucac uugcgagaua cacaacaaag aggcgaaaag 240 aaaagcauga aggcuagcaa gacacgggcu gugcgccuca ccaguuugac ggaggacgug 300 gccgccguac cggaagaugc accauuccgu gcacgccugc acguccucuu cgcccagauc 360 gagcgggagu ucgagcagcu guaucuggag aaccaggccc ugcaggagaa gcuagacaua 420 gccaccacca ccaaggaguc auugauucca cccgaugcca gauccgcagc aggagguuca 480 augagcacuc aggccucauc cgcagcuuug gccacgccug caacccagac ggaugaggug 540 gcgggcagcu uccuggcugc agcauccagu acacacaaga gcuugaaggc caaacuaagc 600 agcgcuaccg gcggcagcaa agccaaggcc agcaacaaga ucaaggccca aaccagccgc 660 auugugucca gcuuuaaggc gcccacagug gucagcacgg ugguccgaga auucggcggc 720 cacaaggaug gcaucuggca gguugccgcc aaggcgggac aacccaucau uggcaccgcc 780 uccgccgauc auacggccug caucuggggc guggagagcg ccaggugccu uuugcaguac 840 caagggcacg ccggcuccgu aaacuccauc aaguuccauc agcagcgaga ucuugugcuc 900 accggcagcg gggaugguac ugcccacauc uggcaggcgg ccgucaacug ggaagugccc 960 aagaagggcc auucuucaga ggaggaacug gacgacagug augagcaggu ggaagaucgc 1020 gaucgugugg acaccuuacg cacuccgcuc ugcgaguuca caggccccgg cggucaucuu 1080 ucagucgugg uggccgccga uuggcucucu ucgauggauc aaaucaucac gggcagcugg 1140 gaucggacug ccauccugug ggacguggag acuggccugc cacugcagcc gcucacuggg 1200 cacgaccacg agcucaccca cgugucggca caucccaccc agcgccuggu ggucaccgcu 1260 uccagggaca cuacguuccg ucuaugggac uuccgagagg caaucccagc ggucuccguc 1320 uuccaagggc acacggaaac gguuacgucc aguguguucg cacgcgacga caagguggug 1380 uccggaucag augaucguac aaucaagguc ugggagcugc gcaacaugcg auccgccuug 1440 gccaccauuc gaacggauuc cucggucaau cguuuggcgg ucuccacugg uggaauuauc 1500 gccauaccgc augacaaucg acagauccga cuguucgauu ugaacgggca gagagucgcc 1560 cgauugccac ggaccagucg gcagggccac cgacggaugg ugucgucugu agcgugggcg 1620 gaggagccgc ugcuggacug cgaucucuuc uccugcggcu ucgaucgccg cguuuucggc 1680 ugguccaucg uucuuccgaa ggacaauuga aggcaacuga agcggagagc ugcugcacca 1740 aaggaaugag gaggcgcagc ucugcgaaau cccaaacauc ucucgaacaa agaagcaucc 1800 ccaaagcgau gcccacauac uuacauauau guaaccguuu aucauagauc aaguauuaug 1860 cagacauugu ucaggauguu cuacuaucuc guugucgccc acugaaugua auuaacguug 1920 cuuuuugcca aauagucgug gcauagcgug cugcacgaac uacauauuau cauuguuaua 1980 gcauacguau guauaugcgu ugggacuggg cuaaacucuu cggagugcag agcuuccgau 2040 guccagaccc gauuuuaucu guguuuuacu accauaauac auauuauaga cgucuaaggc 2100 auuaacgcuu uagccaacau gaauaaacau aaucaaccac ccgauucauc gcuggguaua 2160 uccuaaaa 2168 <210> 34 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> CG12333 sense primer <400> 34 gagctctctg cgaaatccca aacatct 27 <210> 35 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> CG12333 antisense primer <400> 35 ggatcccgct atgccacgac tatttgg 27 <210> 36 <211> 3658 <212> RNA <213> Drosophila melanogaster Dbo mRNA <400> 36 uaugcauuuc ucacaucucu aucgccauac caaucguuuc guguuucgac uuuuccaggc 60 caaccaaaaa acgauuuuuc cguacugaaa agugcaauuc gcgcaaggaa aaucuucgau 120 gugguccuuu uaagccauca agauugcauu uucgaaauuu uccgccugca gcuggcccug 180 gacgugcuuu guauccguag agaacagaga cgcaaagaua gacgccgugu gggguugggu 240 ugcuuccggc ccgcugcgcu uagcagcgaa cagaaugggc gaccugccgg gcucgggcuc 300 caccgcucaa ccacgggaug cugcugucac cgguaccggu gguaauucca cggcuggugg 360 cggcuccucc guuggaucua cggcagugga ccgaccuccg ucgcccgccc gccucucuca 420 cacguccgag aaacauccga aggucacgcu cacugaacua aauaugcuac ggcgccaucg 480 ggagcucugc gauguggugc ucaacguggg cggacggaag aucuuugccc accggguaau 540 ccuguccgcc ugcagcuccu acuucugugc cauguucacu ggcgaauugg aggaaucgcg 600 ccagacugag gucaccauac gcgacaucga ugagaaugcc auggagcugc ucaucgacuu 660 uuguuacacc gcccacauaa ucgucgagga gucgaauguc cagacccucc uucccgccgc 720 cugccugcuu caacugguug agauucagga caucugcugc gaguuccuca aacggcaauu 780 ggaucccacc aacugccugg gcauacgcgc cuuugccgac acacacuccu gccgggaacu 840 ucuacggauc gccgacaaau ucacgcagca caacuuccag gaggugaugg agagcgagga 900 guuucugcug cuccccgucg gccagcuggu ggacaucauu ugcagcgacg agcuaaacgu 960 gcgcuccgag gagcaggucu ucaacgcggu cauguccugg cucaaguaca augucgccga 1020 gcggcgacaa caucuggcac aggugcucca acacguccgu cugccucugc ugucgccaaa 1080 guuccugguu ggcacggugg gcucugaucu ccuggugcgc agcgacgagg ccugccggga 1140 uuugguggau gaggcuaaga acuaucuguu gcuuccgcag gagcgaccgc ugaugcaggg 1200 uccacgcacc agaccgcgaa agccaacucg acgaggcgaa guccuguucg ccgugggcgg 1260 uugguguucc ggcgaugcua uugccuccgu ggagcguuuc gauccucaga ccaacgacug 1320 gaaaaugguc gcucccauga gcaaacgacg cugcggaguu ggcguggccg uguuaaauga 1380 ucuacuuuac gccgugggcg gucacgaugg ccagagcuau cugaauagca ucgagcguua 1440 ugauccacaa acgaaucaau gguccugcga cguugcgccc accacuuccu gccgcaccag 1500 cgucggcgug gcugugcucg acggcuucuu guacgcggug gguggccagg auggcguuca 1560 auguuugaac cacguggagc gcuacgaucc aaaagagaac aaauggucga aaguggcacc 1620 gaugacaaca aggcgacugg gaguggcggu agccgugcuu ggaggauuuc ucuacgcaau 1680 uggcggcucc gauggacagu gcccguugaa caccguagag cgauaugauc ccagacacaa 1740 caaaugggua gcagucagcc cgaugucaac gcgacgcaag caccugggcu gugcuguguu 1800 caauaacuac auuuacgccg ucggcgggcg agacgauugc auggagcucu cguccgccga 1860 gcgcuacaau ccacugacca auaccuggag ccccaucgug gccaugaccu cccgccgcag 1920 ugggguuggc cuggccgucg ucaauggaca gcuguaugcu gugggcggcu uugaugguuc 1980 cgccuauuug aaaaccaucg aggucuacga cccagagacg aaccaauggc guuugugcgg 2040 cugcaugaac uaccgccgac ugggcggcgg cgugggcguu augcgugccc cucagacuga 2100 gaacuauaug uggugcgaaa auaguuuuaa gcaaccgaau uccugaaacu aacucaacuc 2160 ccaucccuuc cauugcugau gaaauguugu cuugucucuc uuuucccccc gaaaacaugc 2220 uaacaaauuc aggauacgga aagauucugu ugucugaugg gaacagcagc cgacaacguu 2280 gcccucgcaa acguugccgc cgccgcacuu cuacuaacaa uauccgcaau cgaagccaaa 2340 ucuaaucgac uaaacaaaac aucuuuucgc cccauugucu acuagcagug uaaucgcuuu 2400 guauuaauug uuaauuguuu auuugguaga gccuaaagcg cauguuuguu agacuuuucc 2460 ggaauaguau uagaguuagc auuguuccca cguucaccug caauaacaca uccccaucac 2520 uccugugcgu aauuauauau gauucuacug uauguuaagc aaguauauuu uauaaauaac 2580 uuaauaucua aauguugccg gugguauuuc auggaaaucg cuagcucaua accaugccuu 2640 cuuucgaaaa aaacaaaaca aaaucaaaua uuaaccuaga aacgaauucg auuaguugua 2700 aauccuuaaa cgaagagcuc uaaagucaag uaacaaauug guauuaaaau aauuuagauu 2760 uaaggaguga auucaagaaa guuuguccga aauuaaacau cacaauggug caauauuucg 2820 aucgaccaac uaaacgcugu uguuuaguca aguacacuuu aaaucuuugc aaacgucuuc 2880 aauaauacca cgacuauaug aagugaaccg aauaauugca aagaauuuaa aaaaauaaug 2940 aauauucagc cugucaagcu uaaaaaaaaa ccauuaaggu aaucacacac acacacaaug 3000 aagaaguuag accuagccca agauaauuca auaguaacaa uuaacaauua cgcagccgua 3060 agaucaacuu uaagguaaua aauuuuaaua uuguuaaucc uagaucgugu aaaccgguaa 3120 gaguuuucgg ccggaaucag gucaaguuuu aagaugacuu auuguuaaac caaguuugac 3180 caucgagcug aaugaaccaa cucgaauuaa ucgcuuagau aagggaguua aaacacaagu 3240 agacucucaa uaacaacaca uuuucuuugu ugccuuuuac accaacacua uuaaacaauu 3300 auuuauucaa aauuauuauu uauuacacgc uuucguaucu cauuuaguuu auuuuaggug 3360 cacauuuaua aacuguuguu aaaucgagca aaaaggaaug aucuuugcuu ccaaaugaug 3420 aauaagaaua gcauuuauaa aaaaccaagc uuauuuauau uauagcuuua agucaaaaug 3480 gucaacaauu auacaaaaau ucgcauacca cccauuaugc guaguuaaaa aauaacauuu 3540 ccuaguugug uugaguggua aaagugcuua acgcaaauaa uagaaguguu aaccuaagaa 3600 augaucguaa acccgugcua uguauaagaa uauauaaaaa uaaaaauaua ucaaaauu 3658 <210> 37 <211> 31 <212> DNA <213> Artificial Sequence <220> <223> dbo sense primer <400> 37 gagctctgtt tatttggtag agcctaaagc g 31 <210> 38 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> dbo antisense primer <400> 38 ggatccccac cggcaacatt tagatat 27 <210> 39 <211> 7606 <212> RNA <213> Drosophila melanogaster Lar mRNA <400> 39 aaagagaaua aaaagagcau uuaaaguuau uaguaaccau cguuguuguu gaguuguugu 60 uguuguuacc aacugucgac auacccuugc aacugccggc gaaaacauag cgaaauaaug 120 ggucugcaga ugacagcagc ccguccaauc gcagcucuca gccuacuagu guuguccuug 180 cucaccugga cucaccccac aaucguggau gcagcccauc cgccggagau caucaggaag 240 ccgcagaauc agggaguccg agugggcggc guugcuagcu ucuauugugc ggcccgcggu 300 gauccgccuc caucgauagu guggcgcaaa aauggcaaaa aaguuucggg aacccagucg 360 cguuacacgg ugcuggagca gcccggcggg auuuccauac uccggauuga gcccgugcgg 420 gcgggacgcg augaugcacc auacgagugu guggcggaga acgggguggg cgaugccguu 480 uccgcagaug caacuuuaac cauauaugaa ggcgauaaaa cacccgcagg cuuuccgguu 540 auaacccagg guccgggaac ucgcgucauu gaaguggguc acacgguccu caugacaugc 600 aaagccaucg gcaauccgac gccaaacauu uacuggauua agaaucagac aaagguugau 660 augagcaauc cgcgcuacuc ucucaaggau ggcuuccugc aaaucgaaaa cagucgcgag 720 gaggaucagg gcaaauacga guguguggcu gagaacucaa ugggcacgga gcacucgaag 780 gccaccaacu uauaugugaa aguccgucgu guuccgccca ccuuuucccg cccaccagag 840 accaucagcg aggugauguu gggaucuaau cugaaucuau ccugcauugc cgucggcuca 900 cccaugccgc augucaagug gaugaagggc uccgaagauc uuacacccga gaaugagaug 960 ccaaucggac gaaauguccu gcagcugauc aauauccagg agagcgccaa cuacacuugc 1020 auagcggccu ccacuuuggg ccaaaucgau uccguuucgg ugguuaaagu gcaaucucug 1080 cccaccgcac ccaccgaugu gcaaaucucc gaggugaccg ccacuucggu gcgucuggag 1140 uggucguaca agggucccga ggacuugcaa uauuacguga uccaguacaa gccgaagaac 1200 gccaaccagg ccuucagcga gauaagcggc aucaucacca uguacuaugu gguccgugcc 1260 cugagucccu acacggagua cgaguucuac gugauagccg ugaacaauau uggacgcgga 1320 ccgcccucgg caccagcgac auguaccacc ggugagacaa aaauggaaag ugcaccacgu 1380 aauguccaag ugcguacgcu gagcucgucc acgaugguua uuacuuggga accaccagag 1440 acgcccaaug gacaagugac cggcuacaag guguacuaca cgaccaauuc gaaucagccg 1500 gaggcgucgu ggaacuccca gauggucgac aauagcgaac ugaccacagu cucggagcug 1560 acgccccacg ccaucuacac gguccggguu caggccuaca caucgauggg agccggucca 1620 auguccacgc cgguccaggu gaaggcccag caaggugugc caucgcaacc gagcaauuuc 1680 cgggcaaccg auaucggcga gaccgcaguc acacugcaau ggaccaagcc gacgcauucc 1740 agcgagaaua ucgugcacua cgagcucuac uggaaugaca cauacgccaa ucaggcccau 1800 cacaagcgca uuuccaacuc ggaggcguau acccuggacg gacucuaccc cgauacccuc 1860 uacuacaucu ggcuggcugc aaggucgcaa cguggcgagg gggccaccac cccgcccauu 1920 ccggugcgca ccaagcaaua uguaccaggu gcuccgccuc gaaauaucac cgccauagcc 1980 accagcucga cgaccauauc ccucagcugg cugccuccgc ccgucgagcg aucgaacggc 2040 cggaucauau acuauaaggu guucuucgug gagguggguc gcgaagacga cgaggccacc 2100 accaugaccc ucaauaugac cagcauugua cuggacgagc ugaagcgcug gacagaguac 2160 aagaucuggg ugcuggccgg caccuccguc ggggaugggc cgcggucgca ucccaucauu 2220 uugcgcaccc aagaggaugu gcccggcgau ccgcaagaug ugaaggccac accuuugaac 2280 uccacuucga uccaugucag cuggaagccg ccucucgaaa aggaucgcaa uggcaucauc 2340 cguggguauc auauacacgc ccaggagcug cgagaugagg gcaagggcuu uuugaacgaa 2400 cccuucaagu uugauguggu agacacgcug gaguucaaug ugacuggcuu gcagccggau 2460 acaaaguacu ccauucaggu ggcggcacua acucguaaag gagaugguga ccggagugcu 2520 gcgauugugg ugaaaacucc uggcggagua ccaguucgac caacggugag ucugaagauc 2580 auggagcggg aaccgaucgu guccaucgaa cuagaauggg agcgaccggc gcagaccuau 2640 ggcgaauugc guggcuaucg acuucgaugg ggcgucaagg accaagcacu gaaggaggag 2700 augcugucag gaccgcagau gaccaagaaa cgguuugaua acuuggaacg cggaguugag 2760 uacgaauuuc guguggcggg cagcaaucau auugguaucg ggcaagagac ggugaaaaua 2820 uuucagacac ccgagggaac acccggugga ccgccuucua acauuaccau ucgcuuccaa 2880 acuccggaug uacugugcgu gaccugggau ccaccaacua gggagcaccg gaauggcaua 2940 aucacccgcu augauguuca guuucacaag aaaaucgauc auggccuggg auccgagcga 3000 aauaugacuc uccggaaggc gguguucaca aaucuggagg agaacaccga guauaucuuc 3060 cgggugaggg cuuauacgaa gcagggagcu ggucccuuca gcgacaaguu aaucguggag 3120 acagaacgug acaugggucg agcaccuaug ucccugcagg cagaggcaac aucggagcaa 3180 acugcggaga uuugguggga accgguaaca agucguggca aguugcuggg cuacaagauc 3240 uuuuacacca ugacagcugu cgaggaucug gacgauuggc aaacgaaaac cguuggacuu 3300 acggaauccg cugaucuugu uaaucucgag aaguuugccc aauaugccgu ggccauugcg 3360 gcgagguuca agaacggauu gggacgucuu agugaaaagg uuacaguacg caucaagccg 3420 gaggaugugc ccuuaaaucu ucgcgcucac gaugucagca cccauucgau gaccuugagu 3480 uggucgccac ccauucgccu aaccccggua aacuacaaga ucagcuucga ugccaugaag 3540 guguuugugg acucgcaggg auucucccag acccagaucg uucccaagcg agagauuauc 3600 cuuaagcacu augugaagac ccacacuauc aacgaacuca guccguuuac cacguacaau 3660 gugaauguga gugccauucc cucggauuau uccuaccggc cgcccacaaa gauuacgguc 3720 acaacgcaaa uggcugcacc ucagccaaug gugaagccgg auuucuacgg cguuguuaau 3780 ggcgaggaaa uucuggugau acugccucag gcuucggagg aauauggacc cauaucgcac 3840 uauuauuugg uggugguccc ggaggacaag uccaaucugc acaagauacc cgaucaguuc 3900 cuuaccgaug aucucuugcc gggcaggaau aagccagagc guccgaaugc accguacauu 3960 gcagccaagu ucccgcagcg uuccauuccg uucacauucc accugggauc uggcgaugau 4020 uaucauaacu uuacaaaucg caaauuggag cgagagaagc gcuaccgcau cuuugugcga 4080 gcgguugugg auacgccaca gaagcaccuc uacaccucca gucccuucuc ugaguuccua 4140 ucgcuggaca ugagggaagc uccgccaggu gagcggcccc accgacccga ucccaauugg 4200 cccgcggagc cggaaguguc ggugaaccgc aacaaggacg aaccggagau ucugugggug 4260 gugcugcccc ugaugguguc cacauucauu guguccaccg cccugaucgu ucucugugug 4320 guuaagcguc gucgccagcc gugcaagacu ccggaucagg cagcugucac aaggccacug 4380 auggccgccg accugggagc cggaccuacg cccagcgauc ccguggacau gaggcgcuug 4440 aacuuccaga cacccggcau gaucucccau ccgcccauac cgauauccga guuugccaac 4500 cacaucgaac gacucaaguc caaugacaau cagaaguuuu cgcaggaaua cgaaagcauu 4560 gagccgggcc aacaguucac cugggacaac uccaaucugg agcauaacaa gucuaagaau 4620 cgcuaugcaa auguuaccgc cuacgaucau ucacgcgucc aguugccagc gguggagggu 4680 gugguuggau cagauuacau caaugccaau uacugugacg gcuaucggaa gcacaaugcc 4740 uacguggcga cccaaggucc guugcaggag accuuugucg acuucuggcg cauguguugg 4800 gaacugaaga cggcaaccau ugugaugaug acgcgauugg aggaacgaac gcgcauaaag 4860 ugcgaucagu auuggcccac ucgcggaacg gaaaccuaug gucagaucuu uguuaccauc 4920 acggagacac aggaacuggc caccuacagc auccgcaccu uccaguugug ccggcagggc 4980 uuuaacgauc ggcgugagau caagcagcug caguucacag ccuggccaga ucauggagug 5040 cccgaucauc cggcgcccuu ccuucaguuc uugcgccggu gucgcgcccu cacgccaccg 5100 gaauccggac ccgugauugu ucacugcucu gcgggaguug gucgcacugg cuguuauauc 5160 guaaucgauu caauguugga acgaaugaag cacgagaaga uuaucgacau cuaugggcau 5220 guuacuuguu uacgggcgca acggaacuac auggugcaga cggaggauca guacaucuuc 5280 auucaugacg ccauccugga ggccaucauc ugugggguga cggaggugcc ggcucgcaau 5340 cuacacaccc accuacagaa acuauugauc acggagcccg gcgagaccau cucgggcaug 5400 gagguggagu ucaagaagcu gucuaacguc aagauggacu cguccaaguu cguaacggcc 5460 aaucugccgu gcaacaagca caagaaucgc cugguccaca uucugccgua cgagucaagu 5520 cgcgucuacc ugacccccau ccauggaauc gagggaagcg acuaugucaa cgccagcuuc 5580 aucgacggcu aucguuaccg uuccgcauac aucgccgcac aggguccugu ccaggaugcc 5640 gcugaggacu uuuggcgcau gcucugggag cacaacucca ccauuguggu caugcugacc 5700 aagcucaagg aaaugggaag ggaaaagugc uuccaguacu ggccucauga gcgauccgua 5760 cgcuaucagu auuaugucgu ggaucccauu gcugaguaca acaugccgca guauaagcug 5820 cgugaauuua aggucacgga ugcccgagau ggcucaucgc gcaccguccg ccaguuccag 5880 uucaucgauu ggccggagca gggugugccc aagucgggcg agggcuucau cgacuucauc 5940 ggacaggugc acaagaccaa ggagcaguuu ggccaggaug gacccauuac cgugcacugu 6000 ucggcgggcg ugggacguuc gggugucuuu aucacucuga gcaucguucu ggaacgaaug 6060 caguacgagg gaguacugga cgucuuccag acagugcgca uacugcgauc ccagcguccg 6120 gcuaugguac aaaccgagga ucaauaccac uucugcuauc gcgcugcacu ggaguacuug 6180 ggcucauucg acaauuauac aaacugagug cauuucucaa ugggauugcc agguccacag 6240 agcuaaaauc ucaucgaccc ugcguucguu gcauacuuau uucgaacucu acgcauaaua 6300 cgcauuuacg auauacacgg auaucauaac ucuaaaguau uauagccagc uacugcccac 6360 acuuacuuaa uacaccuaua cuuauauacg uggauauguu uaguugauaa gcgcaccccc 6420 gcccgcaggu uauguuacca uuacgauucu uugggggacu cgaucuuaua uauugcguuc 6480 uauuuuauug auaauguaaa cuaaagaaga cuguuacacg uuuaauuauc aaaaccuuau 6540 auaucaaccc acaugcacuu cuauguacga guaugugcau caagauuugc ucgcuaaugg 6600 agaaggaugu caaucaacuu guuguuguuc gugguacugc cgccggagug accaugaaug 6660 aggggcgcca cuggaaaccg auacauauuu gaacuaagga cuugguagcu auuagucuua 6720 ccgccuaaau cgaaucucga acggaagcaa agggucgaaa guagaagcca uuuuaaguaa 6780 aucgaaaacu ucuggcgauu uggcgcaaac uccagacgcu auuauaauau auaauacaau 6840 caaucaaaca cugcuauguu aaucaguuuu caugccuugg uuaauggaaa ccaguauaaa 6900 cgguccgugc aauuagucaa aagucuugca uagcgcauau uuuauaaggc agcuagauuc 6960 uaaaauuagu uuucaaacac aacuuacguu uugaucuccc cgcaaaagau gacgaaccuu 7020 auggacugau cgaugucuug guagcuaaca gacaguucaa auuucaaaga ccauuaugau 7080 agaucacaua cgcaacuucc agccaagcag cuauuuuaac uucaaccuca uucccucgac 7140 uucuauucag uucccuagua cauuccugua uauaccacag ucgauacccg agcuagccac 7200 uuauguaugu guaauuuaga uugaaaagcu augaaaaguu auuguaccua agguguauga 7260 agacauagag aauaaccgag cgaacagauu agaaugauaa cuguagauau guacuguagc 7320 auauaauuuu uaccauaaaa uagagcgaca cacgcgguua uaaauaaaca uaaacauaaa 7380 aucgagacau uggcaauagu ucuuaaggua cauaauaaug uauauaacug auaagcuguu 7440 uugcaaaacg uuugaauucg gccucguaac uauacauaaa uauauaaaga guuagcaagc 7500 aaaugcuuaa aagcaaacac uaaauuauua gcugaaacac auguuaaaca accauaguca 7560 uauguaaacu aaauauuuau acgaaauaaa gguauuuguu auaacu 7606 <210> 40 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> Lar sense primer <400> 40 gagctccaat gggattgcca ggtccac 27 <210> 41 <211> 27 <212> DNA <213> Artificial Sequence <220> Lar antisense primer <400> 41 ggatcctccc ccaaagaatc gtaatgg 27 <210> 42 <211> 4507 <212> RNA <213> Homo sapiens FOG2 mRNA <400> 42 ucagcggcag cagccgccgc cgagaugucc cggcgaaagc aaagcaaacc ccggcagauc 60 aaacggccgc uugaagaugc cauugaagau gaggaagaag aauguccauc agaggaaaca 120 gacaucaucu ccaaaggaga cuuuccauug gaggaaagcu uuuccacaga auuugggccu 180 gaaaaucuga gcugcgaaga aguggaauac uuuuguaaca aaggugauga ugaaggaauc 240 caggagacag cagaaucaga uggggacaca cagucagaga aaccggggca accuggaguu 300 gagacagacg acugggaugg accaggagag cuggaggugu uucagaaaga uggggaacga 360 aaaauucaga gucgacagca acuuccagug ggaacaaccu gggggccguu uccugggaag 420 auggacuuga auaauaauuc uuugaagaca aaggcucagg ucccaauggu gcugacugcu 480 ggucccaagu gguugcugga ugugacuugg caaggagugg aagacaacaa aaacaacugc 540 auuguguaca gcaaaggggg ucagcuuugg uguacaacua cgaaggccau cucugagggu 600 gaagagcuaa uugccuuugu gguggauuuu gacucaaggc uacaagcugc cagucagaug 660 acucucacag aagggaugua cccugcacgc cugcuggacu caauucagcu gcuuccucag 720 caagcugcca uggcuucuau uuugcccaca gcuauuguca auaaggauau auucccuugc 780 aaguccugug gcaucuggua ucggagugag cggaaucugc aggcccauuu gauguacuac 840 ugcaguggga ggcaaagaga agcugcuccg gugucagagg aaaaugaaga cagugcccau 900 cagauuucca gccugugccc cuucccacag ugcaccaaga gcuuuucaaa ugcucgagcu 960 cuagaaaugc accugaauuc acacagugga gugaaaaugg aagaauuccu gcccccuggu 1020 gcuagucuaa aaugcaccgu cuguagcuac acugcugauu ccgugaucaa cuuucaccaa 1080 caccuguucu cccaucucac ucaagcugcc uuccgaugua aucacugcca uuucggcuuc 1140 cagacucaga gggaguuauu gcagcaccag gagcuccaug ucccuagcgg caaacuuccc 1200 agagaaagug acauggaaca cucuccaagu gcaacugaag acagcuuaca gccagccaca 1260 gacuuauuga ccagaagcga acuuccccag agccaaaagg ccaugcagac uaaagaugcg 1320 agcucugaca cagagcugga caagugugag aaaaagacuc agcucuuucu cacgaaccag 1380 agaccagaga uacagccuac aacaaauaaa caaagcuuuu cuuacacaaa aauaaagucu 1440 gagcccucua gcccaagacu ugccucaucu ccaguucagc cuaauauugg gccuucuuuc 1500 ccugugggcc cuuuccuauc ucaguuuucu uucccccaag auaucaccau ggucccucaa 1560 gcuucagaga ucuuagcuaa gaugucugaa cuggugcauc ggcgacugag gcauggcagu 1620 aguagcuacc cucccgucau uuacagcccu uugaugccca agggggcuac uuguuuugag 1680 uguaacauaa cauucaauaa uuuggauaau uaucuagugc acaaaaagca uuauugcagc 1740 agccgauggc agcagauggc uaagucccca gaguucccua gugugucaga aaagaugccu 1800 gaagcuuuga gucccaacac uggccaaacc uccauaaacc uucucaaccc agcugcucau 1860 ucugcugauc cugagaaucc acuucuucaa acaucuugca ucaauucuuc cacugucuua 1920 gauuuaauug ggccaaaugg gaagggccau gacaaggacu uuuccacuca aacuaagaag 1980 cucuccaccu ccaguaacaa ugaugacaaa auuaauggaa aaccuguuga ugugaaaaau 2040 cccagugucc ccuuagugga uggggaaagu gacccaaaua agacuaccug ugaagcuugc 2100 aacauuaccu ucagccggca cgaaacauac augguccaca aacaguauua cugugcuaca 2160 cgccacgacc cuccacugaa gaggucugcu uccaacaaag ugccugccau gcagagaacc 2220 augcgcacac gcaagcgcag aaagauguau gagaugugcc uaccugagca ggaacaaagg 2280 ccuccacugg uucagcagag auuucuugac guagccaacc ucaauaaucc uuguaccucc 2340 acucaagaac ccacagaagg gcuaggagag ugcuaccacc caagauguga uaucuuucca 2400 ggaauugucu cuaaacacuu ggaaacuucu cugacgauca acaagugugu uccaguuucc 2460 aaaugugaua cuacucauuc caguguuucc ugccuagaga uggacgugcc cauagaucuc 2520 agcaaaaagu guuuaucuca gucugagcgg acgaccacgu cucccaaaag gcugcuggac 2580 uaucacgagu gcacugugug caagaucagu uucaauaagg uagaaaacua ucuggcccac 2640 aagcagaauu ucugcccggu uacugcacau cagcguaaug accuggguca acuggacggc 2700 aaaguguuuc cgaauccaga aagcgaacga aacagcccug augucagcua cgaaagaagc 2760 auaauaaaau gugagaaaaa ugggaauuug aagcagccuu cccccaaugg aaacuuauuu 2820 ucaucccacc uagcaacccc gcaaggcuug aaggucuuua gugaagcugc ucagcucauu 2880 gcuacaaaag aagaaaacag acauuuguuu cuuccacaau gccuuuaccc uggagcaaua 2940 aagaaagcaa aaggagccga ccagcuuucu ccauauuaug gaaucaagcc aagugauuau 3000 auuucugguu cucuugucau ccauaacacu gacaucgagc aaagcagaaa ugcagaaaau 3060 gaaucuccua aaggccaggc uuccucaaau gggugugcug cgcugaagaa agauucucug 3120 ccauuguugc ccaaaaaucg aggaauggua auagugaaug guggacugaa acaagaugag 3180 agaccugcug ccaacccaca gcaagagaac auuucccaga auccucagca cgaagacgac 3240 cacaaaucuc ccucguggau cucugagaac ccauuagcug ccaaugagaa ugucucacca 3300 ggaauucccu cagcagagga acaguugucu aguauagcaa aaggugugaa ugguuccagc 3360 caggcuccaa ccagugggaa auauugccgg cuaugugaua uccaguucaa caaccuuuca 3420 aacuuuauaa cucacaagaa guuuuauugc ucaucacaug cagcagaaca ugucaaauga 3480 acuaacuaaa caucagucac cuuugguauc aguguuuagu auguuguucu aaccagucca 3540 gaaaaaaaaa uaagcuguuu gaauuacauc ugggcaauca ggagauaauu cauuauggcu 3600 gaguugaaga cuuaaggugu aauuucauua caguccauua guaaagugua uuauuggugc 3660 cauuuucaaa aaaauuaauu uauuuuacca gcaguauuca uagcuguggu uauguuauuu 3720 uuuauuuaaa aacuuuauau uaaagucauu uguaauguua uuguauaguu auuguguagc 3780 acauaugguu ugcacuguau aguagcuuuu aaagaaaaua gucacaauac agaaaagcau 3840 uuuagaaaua gcuucaaaag cacuugugua ucuugauuuu uucuuauaug cuguugcaga 3900 uauauguaua ugcuaaaaua uaacuugcaa agauguucua aauacacaug cuauaaguuc 3960 gccuuaagau uucaauucuu ggauaaucag gcucuguuug cacuuuauau uuuagcagau 4020 acagucucuu agucacuagg cuuugcauuu guauguagcu guauguuucc guccauuuuc 4080 uuaauccuga accuguaugu uaaaugaaga uggcaauuuu uuucuuguau aguacuugua 4140 uuuucuuucg cugaugcagc ucugucucaa uuuuuaaacc uuugcuguua aaugcaauac 4200 uuuauaaaga augaacaaaa uuacuggaag caguauugua aguaaugagg uaguauuaau 4260 caguuuuauc uuuugaaagg cacagucuaa aucgaaaccc uaaacucaau gcugcaagua 4320 ugaauuuaau ucauauauaa gaucuauuua aauauaagag uagcaauacu gcaccuggug 4380 aucacaaaga uaauguucua cuucugauag aaauaauuuc ucaacaaaug uuguuacuau 4440 gcauguauau ggauggaaua aaauuccaga uuguuggaga aaaaaaaaaa aaaaaaaaaa 4500 aaaaaaa 4507 <210> 43 <211> 34 <212> DNA <213> Artificial Sequence <220> <223> FOG2 sense primer <400> 43 gctctagaca gtcacctttg gtatcagtgt ttag 34 <210> 44 <211> 33 <212> DNA <213> Artificial Sequence <220> 223 FOG2 antisense primer <400> 44 cggaattcca tacttgcagc attgagttta ggg 33 <210> 45 <211> 2534 <212> RNA <213> Homo sapiens SEPT7 mRNA <400> 45 agccucgucu gagggggcgg gggacggagg agggagcggg agucgagcga gagccugugg 60 aggaguccgc cugcuguagc gugcguaagc aaggcagcua cgccgggcgg cuacgcugcg 120 gaaucggcgu aggcgccuuu ggagaaucgg cgggcugcgc uccgcugggg cuggucgcgg 180 agggggggag gggaugucgg ucagugcgag auccgcugcu gcugaggaga ggagcgucaa 240 cagcagcacc augguagcuc aacagaagaa ccuugaaggc uaugugggau uugccaaucu 300 cccaaaucaa guauacagaa aaucggugaa gagagguuuu gaauucacgc uuaugguagu 360 gggugaaucu ggauugggaa agucgacauu aaucaacuca uuauuccuca cagauuugua 420 uucuccagag uauccagguc cuucucauag aauuaaaaag acuguacagg uggaacaauc 480 caaaguuuua aucaaagaag gugguguuca guugcugcuc acaauaguug auaccccagg 540 auuuggagau gcaguggaua auaguaauug cuggcagccu guuaucgacu acauugauag 600 uaaauuugag gacuaccuaa augcagaauc acgagugaac agacgucaga ugccugauaa 660 cagggugcag uguuguuuau acuucauugc uccuucagga cauggacuua aaccauugga 720 uauugaguuu augaagcguu ugcaugaaaa agugaauauc aucccacuua uugccaaagc 780 agacacacuc acaccagagg aaugccaaca guuuaaaaaa cagauaauga aagaaaucca 840 agaacauaaa auuaaaauau acgaauuucc agaaacagau gaugaagaag aaaauaaacu 900 uguuaaaaag auaaaggacc guuuaccucu ugcuguggua gguaguaaua cuaucauuga 960 aguuaauggc aaaaggguca gaggaaggca guauccuugg gguguugcug aaguugaaaa 1020 uggugaacau ugugauuuua caauccuaag aaauauguug auaagaacac acaugcagga 1080 cuugaaagau guuacuaaua auguccacua ugagaacuac agaagcagaa aacuugcagc 1140 ugugacuuau aauggaguug auaacaacaa gaauaaaggg cagcugacua agagcccucu 1200 ggcacaaaug gaagaagaaa gaagggagca uguagcuaaa augaagaaga uggagaugga 1260 gauggagcag guguuugaga ugaaggucaa agaaaaaguu caaaaacuga aggacucuga 1320 agcugagcuc cagcggcgcc augagcaaau gaaaaagaau uuggaagcac agcacaaaga 1380 auuggaggaa aaacgucguc aguucgagga ugagaaagca aacugggaag cucaacaacg 1440 uauuuuagaa caacagaacu cuucaagaac cuuggaaaag aacaagaaga aagggaagau 1500 cuuuuaaacu cucuauugac caccaguuaa cguauuaguu gccaauaugc cagcuuggac 1560 aucaguguuu guuggauccg uuugaccaau uugcaccagu uuuauccaua augauggauu 1620 uaacagcaug acaaaaauua uuuuuuuuuu uguucuugau ggagauuaag augccuugaa 1680 uugucuaggg uguucuguac uuagaaagua agagcucuaa guaccuuucc uacauuuucu 1740 uuuuuuauua aacagauauc uucaguuuaa ugcaagagaa cauuuuacug uuguacaauc 1800 auguucuggu gguuugauug uuuacaggau auuccaaaau aaaaggacuc uggaagauuu 1860 ucauugagga uaaauugcca uaauaugaug caaacugugc uucucuauga uaauuacaau 1920 acaaagguuc cauucagugc agcauauaca auaauguaau uuagucuaac acaguugacc 1980 cuauuuuuug acacuuccau uguuuaaaaa uacacaugga aaaaaaaaaa acccuauaug 2040 cuuacugugc accuagagcu uuuuuauaac aacgucuuuu uguuuguuug uuuuggauuc 2100 uuuaaauaua uauuauucuc auuuagugcc cucuuuagcc agaaucucau uacugcuuca 2160 uuuuuguaau aacauuuaau uuagauauuu uccauauauu ggcacugcua aaauagaaua 2220 uagcaucuuu cauaugguag gaaccaacaa ggaaacuuuc cuuuaacucc cuuuuuacac 2280 uuuaugguaa guagcagggg gggaaaugca uuuauagauc auuucuaggc aaaauuguga 2340 agcuaaugac caaccuguuu cuaccuauau gcagucucuu uauuuuacua gaaaugggaa 2400 ucauggccuc uugaagagaa aaaagucacc auucugcauu uagcuguauu cauauauugc 2460 auuucuguau uuuuuguuug uauuguaaaa aauucacaua auaaacgaug uugugaugua 2520 aaaaaaaaaa aaaa 2534 <210> 46 <211> 34 <212> DNA <213> Artificial Sequence <220> <223> SEPT7 sense primer <400> 46 gctctagaga ccaccagtta acgtattagt tgcc 34 <210> 47 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> SEPT7 antisense primer <400> 47 cggaattcga ggccatgatt cccatttcta gt 32 <210> 48 <211> 4078 <212> RNA <213> Homo sapiens RIN2 mRNA <400> 48 cucauugaca caauugccuc ggagaucgga gaacugaaac aggagauggu gcggacagau 60 gucaaccugg aaaauggccu ggaacccgcu gaaacccaca gcaugguaag acacaaggau 120 gguggcuauu ccgaggaaga ggacgugaag accugugccc gggacucagg cuaugacagc 180 cucuccaaca ggcucagcau cuuggaccgg cuccuccaca cccaccccau auggcugcag 240 cugagucuga gugaggagga ggcagcagag guccugcagg cccagccucc ggggaucuuc 300 cugguucaua aaucuaccaa gaugcagaag aaaguccucu cccuccgccu gcccugugaa 360 uuuggggccc cacucaagga auuugccaua aaggaaagca cauacaggau guucuaccau 420 uuaccuugaa guugccuuau gccauuucaa cagccaaguc ggaggcucag cuugaagaac 480 uggcccagau gggacuaaau uucuggagcu ccccagcuga cagcaaaccc ccgaaccuuc 540 caccucccca uaggccucuu uccuccgacg gugucugucc ugccucccug cgucagcucu 600 gccuuauaaa uggagugcau ucuaucaaaa ccaggacgcc uucagagcug gagugcagcc 660 agaccaacgg ggcccugugc uuuauuaauc cccuuuucuu gaaagugcac agccaggacc 720 ucaguggagg ccugaaacgg ccgagcacaa ggacucccaa cgcgaauggc acggagcgga 780 cucggucccc cccacccagg cccccgccac ccgcuauuaa uagucuccac acaagcccuc 840 ggcuggccag gacugaaacc cagacgagca ugccagaaac agucaaccau aacaaacaug 900 ggaacguagc ucugccugga acgaaaccaa cucccauccc uccaccccgg cugaagaagc 960 aggcuucuuu ucuggaagca gagggcggug caaagaccuu gagcggcggc cggccgggcg 1020 caggcccgga gcuggagcug ggcacagcug gcagcccagg uggggccccg ccugaggccg 1080 ccccggggga uugcacaagg gccccgccgc ccagcucuga aucacggccc ccgugccaug 1140 gaggccggca gcggcugagc gacaugagca uuucuacuuc cuccuccgac ucgcuggagu 1200 ucgaccggag caugccucug uuuggcuacg aggcggacac caacagcagc cuggaggacu 1260 acgaggggga aagugaccaa gagaccaugg cgccccccau caaguccaaa aagaaaagga 1320 gcagcuccuu cgugcugccc aagcucguca agucccagcu gcagaaggug agcggggugu 1380 ucagcuccuu caugaccccg gagaagcgga ugguccgcag gaucgccgag cuuucccggg 1440 acaaaugcac cuacuucggg ugcuuagugc aggacuacgu gagcuuccug caggagaaca 1500 aggagugcca cguguccagc accgacaugc ugcagaccau ccggcaguuc augacccagg 1560 ucaagaacua uuugucucag agcucggagc uggacccccc caucgagucg cugaucccug 1620 aagaccaaau agauguggug cuggaaaaag ccaugcacaa gugcaucuug aagccccuca 1680 aggggcacgu ggaggccaug cugaaggacu uucacauggc cgauggcuca uggaagcaac 1740 ucaaggagaa ccugcagcuu gugcggcaga ggaauccgca ggagcugggg gucuucgccc 1800 cgaccccuga uuuuguggau guggagaaaa ucaaagucaa guucaugacc augcagaaga 1860 uguauucgcc ggaaaagaag gucaugcugc ugcugcgggu cugcaagcuc auuuacacgg 1920 ucauggagaa caacucaggg aggauguaug gcgcugauga cuucuugcca guccugaccu 1980 augucauagc ccagugugac augcuugaau uggacacuga aaucgaguac augauggagc 2040 uccuagaccc aucgcuguua cauggagaag gaggcuauua cuugacaagc gcauauggag 2100 cacuuucucu gauaaagaau uuccaagaag aacaagcagc gcgacugcuc agcucagaaa 2160 ccagagacac ccugaggcag uggcacaaac ggagaaccac caaccggacc auccccucug 2220 uggacgacuu ccagaauuac cuccgaguug cauuucagga ggucaacagu gguugcacag 2280 gaaagacccu ccuugugaga ccuuacauca ccacugagga ugugugucag aucugcgcug 2340 agaaguucaa ggugggggac ccugaggagu acagccucuu ucucuucguu gacgagacau 2400 ggcagcagcu ggcagaggac acuuacccuc aaaaaaucaa ggcggagcug cacagccgac 2460 cacagcccca caucuuccac uuugucuaca aacgcaucaa gaacgauccu uauggcauca 2520 uuuuccagaa cggggaagaa gaccucacca ccuccuagaa gacaggcggg acuucccagu 2580 ggugcaucca aaggggagcu ggaagccuug ccuucccgcu ucuacaugcu ugagcuugaa 2640 aagcagucac cuccucgggg accccucagu guagugacua agccauccac aggccaacuc 2700 ggccaagggc aacuuuagcc acgcaaggua gcugagguuu gugaaacagu aggauucucu 2760 uuuggcaaug gagaauugca ucugaugguu caaguguccu gagauuguuu gcuaccuacc 2820 cccagucagg uucuagguug gcuuacaggu auguauaugu gcagaagaaa cacuuaagau 2880 acaaguucuu uugaauucaa cagcagaugc uugcgaugca gugcgucagg ugauucucac 2940 uccuguggau ggcuucaucc cugccuuccu uccuuucuuu uuccuuuuuu uuuuuuuuuu 3000 uuuuuuuuuu acaaagagcc uucauguuuu uauauauuuc auagaaauuu uuauagcagu 3060 ugcagguaaa cugucaggau ugguuuuaaa auauuuuugu aacuuuaaaa uauucuauaa 3120 uuaugcaugu gauuuuaaca uuuaauauuc aaaaauaaau cucuugcugg auuugagagu 3180 auugcauuuu uaaagucucu cuucuguaac uggauguuuu ggcaacuuug uggggagaga 3240 cugcuggauu ucuuaaagca acguauuccu gacacuggcc acagaaugcc uuuggaaauc 3300 ggauguacug uucucuuguu cacguuuagu gguguuuugc uguuuuguuu uuuaaacaaa 3360 ugaugcugag aauaaggaga gaaaugaaug uagagagagg uagagagaga aauaugaacu 3420 cuaacaaagg acugaggagu gcagucugcu gguucaggcu cuucaaaaga uguagaaaaa 3480 gagauagaag gaaccaccua ugcuuaaaau acuguaaaua ugcagugagg uuuggcaaaa 3540 ucuauuccau gugugauuug cuuguagaaa caauuuugaa agccccuuga ggaaaauaaa 3600 aaucaagaag aacacuuuuc ucccuuuucc auacaaauua aaacuuaaca gcaucaaauu 3660 auugggacca gaaaccaagu aauguauaau guggcuuuug uugaguuaaa uaagaugcua 3720 uauaauggag aagaauuuga aaaugcacaa aaaaaucaau cuacauuauc agaaccugca 3780 gugaaauuaa acuuauguua aauaaaacca guuugcaggu gcacaaacua ugagggucuu 3840 guauccacgu aacacaggua guuacaaaaa cauguuauug uacuguguaa agaugcauag 3900 ucaucucauu ugguuggcuu uguaccuugu accuuuuuua gccuuggcuu uuguugaacu 3960 agaacccuca gcacauacug uguuguacuu uuguaaauga uuuuuuaaau ggaauuuugc 4020 acauaauaca uuguaauacu guaugauaau caugugugaa aauaauuuuu gaaauauc 4078 <210> 49 <211> 31 <212> DNA <213> Artificial Sequence <220> <223> RIN2 sense primer <400> 49 gctctagact tcccagtggt gcatccaaag g 31 <210> 50 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> RIN2 antisense primer <400> 50 cataagttta atttcactgc aggttctg 28 <210> 51 <211> 4237 <212> RNA <213> Homo sapiens SHC1 mRNA <400> 51 cccuguagcc gcccgcucag cucuauccug gggaggagua accugaaauu ugcuggaaug 60 ccaaucacuc ucaccgucuc caccagcagc cucaaccuca uggccgcaga cugcaaacag 120 aucaucgcca accaccacau gcaaucuauc ucauuugcau ccggcgggga uccggacaca 180 gccgaguaug ucgccuaugu ugccaaagac ccugugaauc agagagccug ccacauucug 240 gagugucccg aagggcuugc ccaggauguc aucagcacca uuggccaggc cuucgaguug 300 cgcuucaaac aauaccucag gaacccaccc aaacugguca ccccucauga caggauggcu 360 ggcuuugaug gcucagcaug ggaugaggag gaggaagagc caccugacca ucaguacuau 420 aaugacuucc cggggaagga accccccuug gggggggugg uagacaugag gcuucgggaa 480 ggagccgcuc caggggcugc ucgacccacu gcacccaaug cccagacccc cagccacuug 540 ggagcuacau ugccuguagg acagccuguu gggggagauc cagaaguccg caaacagaug 600 ccaccuccac cacccugucc aggcagagag cuuuuugaug aucccuccua ugucaacguc 660 cagaaccuag acaaggcccg gcaagcagug gguggugcug ggccccccaa uccugcuauc 720 aauggcagug caccccggga ccuguuugac augaagcccu ucgaagaugc ucuucgcgug 780 ccuccaccuc cccagucggu guccauggcu gagcagcucc gaggggagcc cugguuccau 840 gggaagcuga gccggcggga ggcugaggca cugcugcagc ucaaugggga cuuccuggua 900 cgggagagca cgaccacacc uggccaguau gugcucacug gcuugcagag ugggcagccu 960 aagcauuugc uacuggugga cccugagggu guggugagug uggcagaggu gugggugagg 1020 gguggcaaaa aggaagguac aguacccuac aguguauccc ucuuccuucc cucauugcuu 1080 uguagacauu cccgcugggc agcugucuau gggccucucu ugcucuuuaa agugucuggc 1140 aucuuccucc agaggcucau cccuuagccu gaacuauguu cuuuauaguc cuggcacagg 1200 gccaggcgug guggcucaca ccuguaaucc cagcgcuuug ggaggcugag gcaggugaau 1260 caccugaggu caggaguucg agaccagccu gaccaauaug gugaaacccu gucucuacua 1320 aaaauacaaa aauuagccgg gugugguggc aggcaccugu agucccagcu acucaggagg 1380 cugaggcagg agaauugcuu gaaucuggga ugcagagguu gcaaugagcu gagaucacac 1440 cacugcacuc cagccugggu gacagaguga gacucugucu gaaaaaacaa aacaaaacaa 1500 auacaguccu ggcccguccu caaaucccca agccuuaagc uuaucagagc uagaaggcag 1560 ucaucucauc ugugccccuc uuuugacaaa uaaggaaacc acccugaaga gguugugaug 1620 cccaauguca gaugguuaga ggcuagacac ugcaaguuuc cuuuuguaag aagauccaag 1680 ggcuucucca gcuccccaga cucaggacuc agaccacugu gcuuucucuu aucugcacug 1740 ccucacuguu cuucaaaacu uuccuuguuu agccaggugu ggugguucau gccuguaauc 1800 ccaacauuuu gcgaagcuga ggugggagga uugcuugagg ccaggaguuc aagaccagcc 1860 ugggcaacau agcgagaccu ugucucuagu caggaauggu gacacaugcc uguaguccua 1920 ccuacccagg aggcugaggc cugaggaucg cuugagccca ggaguucgag guuacaguga 1980 gccaugauug cgccacugua cucuagccug ggugacagag acccugucug gaaacaaaaa 2040 aacuuuuguu uguuuucuac aguaaucuca gcuuccuuuc uuuuuuuuug agauaggguu 2100 uugcuuuguc acucaggcug cgguguggug gcacaccugg cuaauuuuug uauuuuuuuu 2160 uugguagaga ugggguuucg ccauguugcc caagcuguuc uugaacuccc gggcucaagu 2220 gauccgccuc ccucggccuc ccagagugcu gggauuacag gcacggacca ccaugcccag 2280 ccuccacauc uuuuuuugca cuguguauac ucuucugaga caugccaacu uccuccaggu 2340 caagaaaggg guauauagcu cucagcuuca cucuuucagg gcugaugucg ccuuugccuu 2400 uucucacuuc acugaccugu cuauuccuac aacugucucu uucuagagaa gccucaauga 2460 ucaggauuga caggccacac ucucccccac cauuuuuuuc uccuccuuca agccucuugu 2520 cuguuucacc cucuuccacc uuggaggcug aggucuuauu ugacucuuca ccugaauuga 2580 ccuucuuccu ucccacccuc ccagguucgg acuaaggauc accgcuuuga aagugucagu 2640 caccuuauca gcuaccacau ggacaaucac uugcccauca ucucugcggg cagcgaacug 2700 ugucuacagc aaccugugga gcggaaacug ugaucugccc uagcgcucuc uuccagaaga 2760 ugcccuccaa uccuuuccac ccuauucccu aacucucggg accucguuug ggaguguucu 2820 gugggcuugg ccuuguguca gagcugggag uagcauggac ucuggguuuc auauccagcu 2880 gagugagagg guuugaguca aaagccuggg ugagaauccu gccucucccc aaacauuaau 2940 caccaaagua uuaauguaca gaguggcccc ucaccugggc cuuuccugug ccaaccugau 3000 gccccuuccc caagaaggug agugcuuguc auggaaaaug uccuguggug acaggcccag 3060 uggaacaguc acccuucugg gcaaggggga acaaaucaca ccucugggcu ucaggguauc 3120 ccagaccccu cucaacaccc gcccccccca uguuuaaacu uugugccuuu gaccaucucu 3180 uaggucuaau gauauuuuau gcaaacaguu cuuggacccc ugaauucaau gacagggaug 3240 ccaacaccuu cuuggcuucu gggaccugug uucuugcuga gcacccucuc cgguuugggu 3300 ugggauaaca gaggcaggag uggcagcugu ccccucuccc uggggauaug caacccuuag 3360 agauugcccc agagccccac ucccggccag gcgggagaug gaccccuccc uugcucagug 3420 ccuccuggcc ggggccccuc accccaaggg gucuguauau acauuucaua aggccugccc 3480 ucccauguug caugccuaug uacucuacgc caaagugcag cccuuccucc ugaagccucu 3540 gcccugccuc ccuuucuggg agggcggggu gggggugacu gaauuugggc cucuuguaca 3600 guuaacucuc ccagguggau uuuguggagg ugagaaaagg ggcauugaga cuauaaagca 3660 guagacaauc cccacauacc aucuguagag uuggaacugc auucuuuuaa aguuuuauau 3720 gcauauauuu uagggcugua gacuuacuuu ccuauuuucu uuuccauugc uuauucuuga 3780 gcacaaaaug auaaucaauu auuacauuua uacaucaccu uuuugacuuu uccaagcccu 3840 uuuacagcuc uuggcauuuu ccucgccuag gccugugagg uaacugggau cgcaccuuuu 3900 auaccagaga ccugaggcag augaaauuua uuuccaucua ggacuagaaa aacuuggguc 3960 ucuuaccgcg agacugagag gcagaaguca gcccgaaugc cugucaguuu cauggagggg 4020 aaacgcaaaa ccugcaguuc cugaguaccu ucuacaggcc cggcccagcc uaggcccggg 4080 guggccacac cacagcaagc cggccccccc ucuuuuggcc uuguggauaa gggagaguug 4140 accguuuuca uccuggccuc cuuuugcugu uuggauguuu ccacgggucu cacuuauacc 4200 aaagggaaaa cucuucauua aaguccguau uucuucu 4237 <210> 52 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> SHC1 sense primer <400> 52 gctctagact agcgctctct tccagaagat gc 32 <210> 53 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> SHC1 antisense primer <400> 53 gtactcagga actgcaggtt ttgc 24 <210> 54 <211> 1830 <212> RNA <213> Homo sapiens HOXB5 mRNA <400> 54 gugaagcaca ggguuauaac gaccacgauc cacaaaucaa gcccuccaaa aucacccaaa 60 ugagcucgua cuuuguaaac uccuucucgg ggcguuaucc aaauggcccg gacuaucagu 120 ugcuaaauua uggcaguggc agcucucuga gcggcucuua cagggauccc gcugccaugc 180 acaccggcuc uuacggcuac aauuacaaug ggauggaccu cagcgucaac cgcuccucgg 240 ccuccuccag ccacuuuggg gcggugggcg agagcucgcg cgccuucccc gcgcccgccc 300 aggagccccg cuucaggcaa gcggcuucga gcugcucccu guccucgccc gagucccugc 360 ccugcaccaa cggcgacagc cacggcgcca agcccucugc uucguccccc uccgaccagg 420 cgaccucagc cagcuccagc gccaauuuca ccgaaauaga cgaggccagc gcguccucgg 480 agccugagga agcggcaagc cagcuaagca gccccagccu agcucgggcg cagccagagc 540 ccauggccac cuccacagcc gcgcccgagg ggcagacucc gcaaauauuc cccuggauga 600 ggaagcuuca caucagccau gauaugaccg ggccggacgg gaaaagggcc cggaccgcgu 660 auacccgcua ccagacccug gagcuggaaa aggaguucca cuucaaccgc uaccugaccc 720 ggcgacggcg caucgagauc gcccacgcac ucugccuguc cgagcgccag aucaagaucu 780 gguuccagaa ccggcgcaug aaguggaaga aggacaacaa auugaaaagu augagccugg 840 cuacagcugg cagcgccuuc cagcccugag cccgcccaga ggagcccagc ggcccaagag 900 cccgugccac ccccagcccu ggccccucca auccuccccg cucugccgcc gcccgcuggg 960 gaccgguucc cacaagccug ccucgccuug uguuacgaua uuucguuugg ucuuaggucu 1020 uccuguggcu cccucucucc uggacugguu aucuuguuau uauuguuaau aauaauuauu 1080 auuauuauuu uccuuccaug cucccaacuc ccuucugcuu gucccaaauc cgccaguguu 1140 ucugaauguu ugugucugug guugcagucu uucccccagg aaaaaaaaaa aaagaaauuc 1200 gcauguuuaa ugugaacucu ccccucccca ucuguguucu aacuuauuua uaaaaagaug 1260 aucgcuguau uuugaguuuc agcuggaaac uucuguaagg ggcagcaguu gagguggggu 1320 agugccgcag uggggucaag cugagcuggc uucggagaug gagucccuuu ucauucuccu 1380 ccuccucccu ccucacuccc uaggcccaag ucuccuaggg gcuugguccu agggugggaa 1440 ggggcuaggg aggaccaaag ggaugguauu gagaagagag aaagaagaua gugagauuua 1500 aguuccugcu gccuggguag gccccacaag gccuggucug ggaguauacg gaaacaaaaa 1560 ugauccucag ugcaaaaugu cuuguguauu ucucugugaa uccauggguc uggcuagagg 1620 gcccaaagcu uguaaauaug gggauagucu gggucagacc caucucuccc uuacccaucu 1680 ugcuuccaag accauuugua gugagcgagu ggaugcugug cuacguguga aaucugucuu 1740 ugcggggccu gucucaguga uucgcuuuug guauuuguuu guagcuuucc uggaagucaa 1800 Auaaauguuu cccccacucc aaaaaaaaaa 1830 <210> 55 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> HOXB5 sense primer <400> 55 gctctagaca gaggagccca gcggccca 28 <210> 56 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> HOXB5 antisense primer <400> 56 cggaattcca ctgagacagg ccccgcaaag aca 33 <210> 57 <211> 4051 <212> RNA <213> Homo sapiens KLF11 mRNA <400> 57 gagggccgcg ccggggcaga gccgcgcggg cgggcgaggc gcgugccggc cgcaggagcu 60 ccggguugcc gccgccgccg ccgcccgcag cccacgugcg gccgcugcug cgcccgagcu 120 cacgccccgc ggccgcuuug uugcucccgg ccggccugca cgaugcacac gccggacuuc 180 gcaggcccag acgacgcgcg cgcaguugac aucauggaca uaugugaguc cauccuggag 240 aggaagcggc augacagcga aaggucuacu ugcagcaucu uggagcagac agacauggaa 300 gcugucgagg cucuuguuug uaugagcucc uggggucaaa gaucccagaa aggugaccug 360 uugcggauaa gaccccucac gccugucucu gacucugggg augucaccac cacugugcau 420 auggaugcag ccacaccuga acuaccaaaa gacuuccauu cuuuaucgac ucugugcaua 480 acuccuccuc agagcccuga ucucguggag ccaucgacaa ggacaccugu uucuccccaa 540 guaacagauu ccaaagcaug uacagccacg gauguucucc aguccucugc cguaguggcc 600 agagcucuga gcgggggcgc ggagaggggc uugcuggguu uggagccagu gcccagcucu 660 cccugcaggg ccaaggggac uagcgugauc cgacacacug gggagagccc ugcugccugc 720 uuucccacca uccagacucc agauugccgg cuuucugaca gcagagaagg agaagagcag 780 cuucugggac acuuugaaac uuugcaggac acacaccuca cggacaguuu acucagcacu 840 aacuuggugu ccugucagcc cugcuugcac aagucuggug gccugcugcu cacugacaaa 900 ggccagcagg caggguggcc uggugcaguu cagacuugcu caccaaagaa uuaugaaaau 960 gaccugccca ggaaaaccac cccucugauu ucugucucug ucccugcucc cccuguccuu 1020 ugccagauga ucccugugac uggacaaagu agcauguuac cagcuuuuuu gaagcccccu 1080 ccccaguugu cuguggggac ugugagaccc auccuagcuc aggcugcucc agcgccucaa 1140 ccuguguucg ugggaccugc ugugccucag ggagcuguga uguugguccu gccccaggga 1200 gcccucccuc cgccugcccc cugugcagcc aaugucaugg cugccgggaa uaccaaguug 1260 uugccccuug ccccugcucc aguguucauc accucuagcc aaaacugugu cccucaggua 1320 gacuuuuccc gaaggaggaa cuauguuugc agcuucccag guugccggaa gaccuacuuc 1380 aaaaguuccc accuuaaggc ccaucuucgc acucacacag gggagaagcc uuucaacugc 1440 agcugggaug gcugugauaa aaaguuugcu cguucggaug agcugucacg ccaccgcaga 1500 acucacacag gggagaagaa guuugugugc ccggugugug accgacguuu caugcgcagu 1560 gaccaccuga cgaagcaugc ccggcgccac augacgacca agaagauccc aggcuggcag 1620 gcagagguug gcaagcugaa cagaaucgcc ucugcagaga gcccggggag cccacuggug 1680 Agcaugccag ccucugccug aaagguccau uaggacauca cucaugggau uuuuaaaaag 1740 ccucuuucca ggaauggaac ugauggauuc cucucccacu gccucaccca aaaaaaacgg 1800 ucuuggcggc cuaggggaag aucggggagc ugguuuugau gaaaguaugu uaacuuuucu 1860 uuuccacuug ggacccuguu caguaucuuu uguaguuuca gaaguuuuuu uguuuugguu 1920 uuuuuuuuaa agaaauggua gaaaauuuga uaaucugaau caccagcauu caaacaaaua 1980 uuucggcaau aaaguuuaca aaaucuggau uuuuacaacc uuuucuauug auguuuugua 2040 gaaauaagac aggguacuaa uuuuuauacu gguuuuuaga aaaauauuua uauuguuggu 2100 gcucaaauca ccaauuucua gcuagaucau uuugcagccu ucuuuucagu guuuaauaac 2160 aaaguuuuuc cuaauggccc uucuuuuagu aaacuggaca uguuauucca cuacaaaaac 2220 cacaaguuau cuggccuuuu agaucuuuuu ggaaucggac cugguugagu aaggaccucu 2280 uaaaagggaa aaauaaauuu ugccgucagc uucuucauaa cguuuucaag gaaauucuag 2340 gcaaucauuc cugucaccaa agaacuaaaa uuuugguuga cuggaacuag ugagcugugu 2400 ccauggugug ucaugaagga uguaccccag agaguaacau gagccacugg gcagauccca 2460 gggaccagua cuugcugcag gaucuagucu guaauagucu uggccauggc ucugcugaaa 2520 gcaagccauu caguuucuug uuuguaccua aaacaccaaa aaagaaacac ucaaauccag 2580 cugcuuuguc aauugucagu ucugacuccu uuugcugugg ccuuauccgu acuauauugu 2640 ggguagagua acuucucaga aaaaaaggaa augucuguau ugguuggaug aaacuccacc 2700 agagcacagc uuagcugggg cgagaugcau gugaaggcag gcagugccaa gauuccgcuu 2760 ccuuuguuug ccaaauacua gaaacacaag gaaaugcaag uuacgcuaaa uggcaguaau 2820 acuacccaac ugccuuucug uucauuuugu uugaaggaaa uuguuuugac caaacagaaa 2880 auuacuugga augguguguu uuacagucua ccuagaaaau agauggacaa uauuuuucaa 2940 cuguaugagc acguagauaa ccgagagaau guggccaccu guguucaaga agccacugau 3000 acugguuuuu guuaaacauu ggaaguucag gcaauggaau aaauguagga acauacagaa 3060 uguugcacua auuugguagc cugggaauuu uuuuuauugu gcaguaugua uuuaaauuuu 3120 gucuauguua auuaccagca uuuaccuuua uuuaaaugau gguaaggugg aauauugaau 3180 aaaauuaggu uuuguguuuu guucuuugua gucugauaaa aucuccaccu ggucauucau 3240 ugugugugac uugauaccug uuaacuugcc ccuuaguauc agcuguuacu ugacacaaau 3300 guguguguua uucagagguu uucagucugg acacuccaua ggugaguguc gugucuucgu 3360 gagacagcac aguugucuca uguugugcau aguucauguu uccucaccac ccaguccuuu 3420 cuccugcuca ucaaaaucag cauacacauu uuugacugua cacacuauaa auggcaucaa 3480 auuuggauau uuuucuuaau uaugacaugc aaaguaaugu gaguccugcc aguauucugg 3540 uggauaaggu cuuuugagua uuugguugcu ugucacaaca uucuccaagc agugauauuu 3600 cuaaagagga gauacauguu gaaaacgguu uuaauuuaca cuuccauuuc cugauuacau 3660 uuggaaauac uuuguguaaa ccaucccccu uccaccucca uuugucuguu gaaagauuuu 3720 aaguuggaaa caguuccugu cugaaaacuc uucugagaac cacaaaccuu guguauggau 3780 ucggcaugga gcccucagcu ggcggcucug ggugcugacg gccgcuggag aggugggcuc 3840 cccucgugca cuuuauugcc ugggcaguuu ugcuugaucu uuugugacuu ugagccuuuu 3900 aaguaguuug aaugauaaga cuuaaaaugu uucauaauua uguuuuaugu aacagacuuu 3960 gacauuauuu aaacgagcau guguaaugua acuuuucucu uugaaucaua uaaaacuuga 4020 uuuuacauug aaaaaaaaaa aaaaaaaaaa a 4051 <210> 58 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> KLF11 sense primer <400> 58 tctagaggtc cattaggaca tcactc 26 <210> 59 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> KLF11 antisense primer <400> 59 gaattcctgc ccaggcaata aagtg 25 <210> 60 <211> 2779 <212> RNA <213> Homo sapiens TCF7L1 mRNA <400> 60 gcgccgggcc gggccgggca gggcgcgggc ggcuaggggc uccgagagcg gcggccccgg 60 cccgcggccc caccaugccc cagcucggcg gcgggggcgg cggcggcggc ggcggcagcg 120 ggggaggcgg cggcuccagc gccggggcgg ccggcggagg ggacgaccuc ggggcgaacg 180 acgagcugau ccccuuccag gacgaggggg gcgaggagca ggagccgagc agcgauagcg 240 ccucggcgca gcgggaccua gacgagguca agucgucccu ggucaacgag ucggagaacc 300 agagcagcag cucggacucg gaggcggaga ggcgcccgca gcccguccgg gacacuuucc 360 agaagccgcg ggacuauuuc gccgaaguga gaaggccuca ggacagcgcg uucuuuaaag 420 gacccccgua cccuggguac cccuuccuga ugaucccgga ccugagcagc ccguaccucu 480 ccaacggacc ccugucuccc ggaggagcgc gcaccuaccu gcagaugaaa uggccccucc 540 ucgauguccc cuccagcgcc acagucaagg acacgagguc accaucucca gcacacuugu 600 cuaauaaagu uccugucguu cagcacccgc aucacaugca uccgcugacu ccccucauca 660 ccuacagcaa ugaccacuuc ucccccggcu ccccucccac ccaccucucc ccagagaucg 720 auccaaagac aggaaucccc cggcccccuc acccauccga gcugucaccg uauuacccac 780 ucucucccgg agcugucgga caaauccccc acccccucgg cuggcucguc ccacagcaag 840 gccagcccau guacucccuu ccucccggug gcuuccggca cccuuacccc gcccucgcca 900 ugaacgccuc gauguccagc cuggucucca gucgguucuc uccucacaug guggcuccug 960 cccacccugg ccugcccacc ucagggaucc cccacccugc caucgucucc cccaucguca 1020 agcaggaacc ggcacccccc agccugagcc cugcagugag cgugaaauca ccagucaccg 1080 ugaaaaagga ggaggaaaag aagccccacg ugaagaagcc ucugaaugcc uucauguugu 1140 auaugaagga gaugagggcc aagguggugg cugagugcac ccugaaggaa agugcagcca 1200 uuaaccagau ccuuggaaga aaguggcaca accugucucg agaagaacag gccaaguacu 1260 acgagcuggc ccggaaggag cggcagcuuc acucgcagcu cuacccaacc uggucagccc 1320 gggacaacua ugguaagaaa aagaagagga agagagaaaa gcagcugucc cagacacagu 1380 cacagcagca aguccaggag gcagagggug cccuggccuc caagagcaag aagccaugug 1440 uucaguaccu gccccccgag aagcccugug acagcccugc cuccucccac gggagcaugc 1500 uggacucccc ggccacuccc ucugcagcuu uggccucacc agcugccccu gcugccaccc 1560 auucggagca agcccagccc cucucccuca ccaccaaacc agaaacccgg gcccagcugg 1620 cucuccacuc ugccgccuuc cugucggcua aggcugcagc cuccuccucu gggcagaugg 1680 gcagccagcc uccccuccug ucccggcccc ucccccuugg guccaugccc acagcucugc 1740 uggccucucc cccguccuuc cccgccacgc uccaugccca ccaggcccuc ccggugcuac 1800 aggcccagcc ucuuucccug gucaccaagu cugcccacua agcucccccc gaccccugca 1860 ggcugucaca ugacucauug aguaguaaug auucagaaga aaaagaaaaa ggagacuuua 1920 uuggucaaua uuugaccacu cuggacuguu cuguaaagug gcugguaaca acagcacuuu 1980 acaguuugua gauguaacca guagcugauc uuaaggcuuu uuuaaaaaac aaaacaaaac 2040 aacaaaaaaa aaucuuuaua agaaagagaa cugaaaagua gcgugcuauu cguccuguag 2100 gugcuguggu ggauggaccu gggcagaggg cacuucucuc ucuuaccucu cuugcacuuu 2160 cugucuccug ucucuucucg ccccugccgc cugccccagc uuccccgacu ccaucugcag 2220 cucugccauu gugacauuuc cuguuaccca gcccaaguuu ucaucgucug cucaauaccg 2280 uggguucuuc uucguccucu guccucugcc cagugugagg ccaucaccau gugagaagac 2340 aucuuggccu gauuugcugc caccagcguc cccucccuca gugggcccga acucgccagc 2400 cccagcuuuc aguggagaaa gcgguccucu gaaaugguuu ccucccaacc cccgcauuua 2460 aagggacuca aggugccugc cacuuccuca gcgaagaagu cuguguuccu ccccguccuu 2520 gccaguggcg aucaucccuu cacaauccca gaguggcagg cgggaccagc cccauggucu 2580 ggcuccuguc accugggucc gugccagcac aaucugccaa aguucuagag acccuguucc 2640 cuuccccauc accucacaug cuucuucugu guguauuucu uuuuguuuuu augguuuuug 2700 gagcaauuua aacucccagu uguuuauuuu cacaaaagaa aauaaaauug caguugcaag 2760 aaaaaaaaaa aaaaaaaaa 2779 <210> 61 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> TCF7L1 sense primer <400> 61 gaattcgacc cctgcaggct gtcac 25 <210> 62 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> TCF7L1 antisense primer <400> 62 gatatcggga gtttaaattg ctcc 24 <210> 63 <211> 6797 <212> RNA <213> Homo sapiens ERBB2IP mRNA <400> 63 aauguuggcu caauuaagaa acaucaggga gauaaauuca acccagugug ucuaaaaaug 60 acuacaaaac gaaguuuguu ugugcgguug guaccauguc gcugucuacg aggggaagag 120 gagacuguca cuacucuuga uuauucucau ugcagcuuag aacaaguucc gaaagagauu 180 uuuacuuuug aaaaaaccuu ggaggaacuc uauuuagaug cuaaucagau ugaagagcuu 240 ccaaagcaac uuuuuaacug ucagucuuua cacaaacuga guuugccaga caaugauuua 300 acaacguuac cagcauccau ugcaaaccuu auuaaucuca gggaacugga ugucagcaag 360 aauggaauac aggaguuucc agaaaauaua aaaaauugua aaguuuugac aauuguggag 420 gccaguguaa acccuauuuc caagcucccu gauggauuuu cucagcuguu aaaccuaacc 480 caguuguauc ugaaugaugc uuuucuugag uucuugccag caaauuuugg cagauuaacu 540 aaacuccaaa uauuagagcu uagagaaaac caguuaaaaa uguugccuaa aacuaugaau 600 agacugaccc agcuggaaag acuggauuug ggaaguaacg aauucacgga agugccugaa 660 guacuugagc aacuaagugg auugaaagag uuuuggaugg augcuaauag acugacuuuu 720 auuccagggu uuauugguag uuugaaacag cucacauauu uggauguuuc uaaaaauaau 780 auugaaaugg uugaagaagg aauuucaaca ugugaaaacc uucaagaccu ccuauuauca 840 agcaauucac uucagcagcu uccugagacu auugguucgu ugaagaauau aacaacgcuu 900 aaaauagaug aaaaccaguu aauguaucug ccagacucua uaggaggguu aauaucagua 960 gaagaacugg auuguaguuu caaugaaguu gaagcuuugc cuucaucuau ugggcagcuu 1020 acuaacuuaa gaacuuuugc ugcugaucau aauuacuuac agcaguugcc cccagagauu 1080 ggaagcugga aaaauauaac ugugcuguuu cuccauucca auaaacuuga gacacuucca 1140 gaggaaaugg gugauaugca aaaauuaaaa gucauuaauu uaagugauaa uagauuaaag 1200 aauuuacccu uuagcuuuac aaagcuacag caauugacag cuauguggcu cucagauaau 1260 caguccaaac cccugauacc ucuucaaaaa gaaacugauu cagagaccca gaaaauggug 1320 cuuaccaacu acauguuccc ucaacagcca aggacugagg auguuauguu uauaucagau 1380 aaugaaaguu uuaacccuuc auugugggag gaacagagga aacagcgggc ucaaguugca 1440 uuugaaugug augaagacaa agaugaaagg gaggcaccuc ccagggaggg aaauuuaaaa 1500 agauauccaa caccauaccc agaugagcuu aagaauaugg ucaaaacugu ucaaaccauu 1560 guacauagau uaaaagauga agagaccaau gaagacucag gaagagauuu gaaaccacau 1620 gaagaucaac aagauauaaa uaaagauacc ucagaaagua cuacuacagu aaaaagcaaa 1680 guugaugaaa gagaaaaaua uaugauagga aacucuguac agaagaucag ugaaccugaa 1740 gcugagauua guccugggag uuuaccagug acugcaaaua ugaaagccuc ugagaacuug 1800 aagcauauug uuaaccauga ugauguuuuu gaggaaucug aagaacuuuc uucugaugaa 1860 gagaugaaaa uggcggagau gcgaccacca uuaauugaaa ccucuauuaa ccagccaaaa 1920 gucguagcac uuaguaauaa caaaaaagau gauacaaagg aaacagauuc uuuaucagau 1980 gaaguuacac acaauagcaa ucagaauaac agcaauuguu cuucuccauc ucggaugucu 2040 gauucaguuu cucuuaauac ugauaguagu caagacaccu cacucugcuc uccagugaaa 2100 caaacucaua uugauauuaa uuccaaaauc aggcaagaag augaaaauuu uaacagccuu 2160 uuacaaaaug gagauauuuu aaacaguuca acagaggaaa aguucaaagc ucaugauaaa 2220 aaagauuuua acuuaccuga auaugauuug aauguugaag agcgauuagu ucuaauugag 2280 aaaaguguug acucaacagc cacagcugau gacacucaca aauuagauca uaucaauaug 2340 aaucuuaaua aacuuauaac uaaugauaca uuucaaccag agaucaugga aagaucaaaa 2400 acacaggaua uugugcuugg aacaagcuuu uuaagcauua auucuaaaga ggaaacugag 2460 cacuuggaaa auggaaacaa guauccuaau uuggaauccg uaaauaaggu aaauggacau 2520 ucugaggaaa cuucccaguc uccuaauagg acugaaccac augacaguga uuguucuguu 2580 gacuuaggua uuuccaaaag cacugaagau cucuccccuc agaaaagugg uccaguugga 2640 ucuguuguga aaucucauag cauaacuaau auggagauug gagggcuaaa aaucuaugau 2700 auucuuagug auaauggacc ucagcagcca aguacaaccg uuaaaaucac aucugcuguu 2760 gauggaaaaa auauagucag gagcaagucu gccacacugu uguaugauca accauugcag 2820 guauuuacug guucuuccuc aucuucugau uuaauaucag gaacaaaggc aauuuucaag 2880 uuugauucaa aucauaaucc cgaagagcca aauauaauaa gaggccccac aaguggccca 2940 caaucugcac cucaaauaua ugguccucca caguauaaua uccaauacag uagcagugcu 3000 gcagucaaag acacuuugug gcacuccaaa caaaaucccc aaauagacca ugccaguuuu 3060 ccuccucagc uccuuccuag aucagagagc acagaaaauc aaaguuaugc uaaacauucu 3120 gccaauauga auuucucuaa ucauaacaau guucgagcua auacugcaua ccauuuacau 3180 cagagacuug gcccagcaag acauggggaa augugggcca ucucaccaaa cgaccgacuu 3240 auuccugcag uaacucgaag uacaauccag cgacaaagua guguguccuc cacagccucu 3300 guaaaucuug gugauccagg cucuacaagg cgggcucaga uuccugaagg agauuauuua 3360 ucauacagag aguuccacuc agcgggaaga acuccuccaa ugaugccagg aucacagaga 3420 ccccuuucug cacgaacaua cagcauagau gguccaaaug caucaagacc ucagagugcu 3480 cgacccucua uuaaugaaau accagagaga acuaugucag uuagugauuu caauuauuca 3540 cggacuaguc cuucaaaaag accaaaugca aggguugguu cugagcauuc uuuauuagau 3600 ccuccaggaa aaaguaaagu uccucgugac uggagagaac aaguacuucg acauauugaa 3660 gccaaaaagu uagaaaagag caugcuguca agguccuuua auuccaauuu uacuacugua 3720 agcaguuuuc acuguggcag cucuagggau cugcauggca gccagggcag ucuugccuug 3780 aguguugcag acagaagagg uucugguggg cacauuuuuc gaaugccuuu gaguaaugga 3840 cagaugggcc agccucucag gccucaggca aauuauaguc aaauacauca ccccccucag 3900 gcaucugugg caaggcaucc cucuagagaa caacuaauug auuacuugau gcugaaagug 3960 gcccaccagc cuccauauac acagccccau uguucuccua gacaaggcca ugaacuggca 4020 aaacaagaga uucgagugag gguugaaaag gauccagaac uuggauuuag cauaucaggu 4080 ggugucgggg guagaggaaa cccauucaga ccugaugaug augguauauu uguaacaagg 4140 guacaaccug aaggaccagc aucaaaauua cugcagccag gugauaaaau uauucaggcu 4200 aauggcuaca guuuuauaaa uauugaacau ggacaagcag uguccuugcu aaaaacuuuc 4260 cagaauacag uugaacucau cauuguacga gaaguuuccu cauaagcacu guggacaaaa 4320 aaagcgggga agacagcaag auuuauugga agauacuuac aggggaaauu aauauuuuga 4380 cuauuuuuau auauaaagaa gaacucaaaa aauuauguuc aaauuuguac auuaaugaaa 4440 uaauggaacu ugugguuaga gggaaagaac cacuguacag aauauaaagg agacuguuga 4500 auucauacca uauaaaacuu guuagguuuu uaaacauagc aaucaaggcu acaaaaacaa 4560 accuguguug uuuuuguaua gauuguaggu uuauuuuugg auuucauaua caugacugaa 4620 cugugugcaa ggcaauaguu agccuugauu uuagcccaga gacagauggc agagcuaucu 4680 cucucauagc uuuuaugccc uuauuuuuau ucaacuggua uuaauguuuu ucuccugaaa 4740 cuacuuuuuu ugaugugggc aagagauuug aaguguuggc uuuugcuaug ugcauauuga 4800 auugaagagu gaguagguga agguggugcu gguggguuca cuuuccaagg ccagacuaaa 4860 acaguuauuu ucuauaaaaa ucuggaagca aagaaugggg auggggagag cuacguggua 4920 guauguuuuu auuaggagaa uaaugcaaua aaauauguaa ugucuuuuuu auaaagcaaa 4980 aaagacaaua auugcauuua ugagcucggc aggaucuguu cuugucauag ccauugacua 5040 uacauuugcu acuggugauu caguuuuuaa uuuuuuaguc acaggaaauu uuuaacucua 5100 cuguagaugc auguccaugc auuuucugug uuauggaaau ccacugauuu uuuuuuuuuu 5160 uucaaauggu gguacuugca aucuguuuua uaauuagugc uccauuuaaa ucuaauuuau 5220 aauuuuuauu uuaagcagca aaugaaacaa aaauggccag uuuuaagauu guguugccug 5280 uaacacaaaa uguuacgaag guuuaggaaa gccucuuuga uuuuuguuug gccuugcauu 5340 gccuugguaa aguaaaagga aacaguacac uuggagcuag gaaaccaaag caagcuuugu 5400 gaaacuggca cagugauaga gaauugcugu ggagaguuau agagcaaagg gauggguccu 5460 ugaggccugc caguguguaa agguguucaa auaaagggcu guuucuacag guaacauuaa 5520 augugaacuc aacacuucca gagucuuuaa aggguuucua uguguaucag uguaauagug 5580 uuuuaccacc aacugccuuu cuuuguuccu aguuacugua acaaauauuu gaugauagag 5640 guuuauuaau uuuguuuauc cagaccauua auuuuauuug uuuuuguucu auguaaucaa 5700 auaaaauuug aguaacaugu aaugguaagg auuaaugcau gguuauuugg accagaaaaa 5760 agugccauag aagaccaaua acuguuuagu ugaggcuagu cuggaaccuu ucauuagagc 5820 aauauuuggu uauugcacuu cauuuuuauu uacuaagaaa ugcaauuugg gaauuuuuaa 5880 ucuguuaugc uuuguuuauc aaccuugauu uuaauuaaga cuuuuauaag acuagcuuaa 5940 aacaccaacc aacauuauuu uugcaaaagu gaguuggacu cacuuuccau ucuugcuagu 6000 cagaguaagu aggcagcacu uuuaaaaaua ugugaacuca aauauugcac uucuuucaag 6060 auguuaucaa uugguuauug uacuguauag uuuuaauaau uuugauugaa acccuuuaac 6120 aacucuuugu aaauuuuaac ucauuuuagu ugauuuucag uacuauuuac auaggaauug 6180 auuuuuaugg auauaguaga agaaaugugc uguauuuuga uaaaauucac uuauuguaug 6240 uguguuguaa ucuaaaaaaa aaaagaauga caaacagcuu cuuuaagaca agucucggug 6300 uucccuuuau ucuuaguuug uuuuuaaaua uuaauuuugg cauucuaaaa uagcuaacau 6360 uucuuuuauu gauuucagau uuucacaggc acauucuacu uuuaaucaga aauauauuua 6420 auaaguauaa uugugaaguu uucaacuacu uuaccuugaa ccacauauac caauuauaau 6480 uuuggaaaag gaauuaagcc ucacggaaca auggaucuuc agcaaaccuu aacuucauug 6540 ucugcacauu acauugaagu auuauaaaug caacagaugu uauaugcacu ggcauuuuau 6600 ccuacucuag uuaguuaaaa uuuuauagua uucuugcaac acauaaaguu gcguaagaaa 6660 cuuuaccaag aggaguauua uagccaaguu uucuuugaaa guauuggaaa acuaaaauua 6720 aaugacaagg acuuugaauu agaauuuugc uguaauaaag uuucaaaauu ugaauaaaau 6780 aauuaaauuu uuugagg 6797 <210> 64 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> ERBB2IP sense primer <400> 64 ggggagagct acgtggtagt atg 23 <210> 65 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> ERBB2IP antisense primer <400> 65 ccaatacttt caaagaaaac tt 22 <210> 66 <211> 3596 <212> RNA <213> Homo sapiens ELMO2 mRNA <400> 66 ucccaagaag gcggggcggg cggaggcugg aggagccgcc gagcggagac ccgggagcag 60 gagcugggcc uaggucugcg cccugaauua gagcccauug ggaacgaugc caccaccguc 120 agacauuguc aaaguggcca uugaguggcc aggugcuaac gcccagcucc uugaaaucga 180 ccagaaacgg ccccuggcau ccauuaucaa ggaaguuugu gauggguggu cguugccaaa 240 cccagaguau uauacccucc guuaugcaga ugguccucag cuguacauca ccgaacagac 300 ucgcagugac auuaagaaug ggacaaucuu acaacuggcu aucuccccgu cccgggcugc 360 acgccagcug auggagagga cccagucauc caacauggag acccggcugg augccaugaa 420 ggagcuggcc aagcucucug ccgacgugac uuucgcuacu gaguucauca acauggaugg 480 caucauugug cugacaaggc ucguggaaag uggaaccaag cucuuguccc acuacaguga 540 gaugcuggca uucacccuga cugccuuccu agagcucaug gaccauggca uugucuccug 600 ggacaugguu ucaaucaccu uuauuaagca gauugcaggg uaugugagcc agcccauggu 660 ggacguguca auccuucaga ggucccuggc cauccuggag agcauggucu ugaacagcca 720 gagucuguac cagaagauag ccgaggaaau caccguggga cagcucaucu cacaccucca 780 ggucuccaac caggagauuc agaccuacgc cauugcacug auuaaugcac uuuuucugaa 840 ggcuccugag gacaaacgac aggauauggc aaaugcauuu gcacagaagc aucuccgguc 900 uauaauccug aaucauguga uccgagggaa ccgccccauc aaaacugaga uggcccauca 960 gcuauauguc cuucaagucc uaaccuuuaa ccuucuggaa gaaaggauga ugaccaagau 1020 ggaccccaau gaccaggcuc aaagggacau cauauuugaa cugaggagga uugcauuuga 1080 cgcagagucu gauccuagca augccccugg gagugggacc gaaaaacgca aagccaugua 1140 cacaaaggac uacaaaaugc ugggauuuac caaccacauc aauccagcca uggacuuuac 1200 ccagacuccu ccuggaaugc uggccuugga caacaugcug uacuuggcua aaguccacca 1260 ggacaccuac auccggauug ucuuggagaa caguagccgg gaagacaaac augaaugccc 1320 cuuuggccgc agugccauug agcucaccaa aaugcucugu gaaauccugc agguugggga 1380 acuaccaaau gaaggacgca augacuacca cccgauguuc uuuacccaug accgagccuu 1440 ugaagagcuc uuuggaaucu gcauccagcu guugaacaag accuggaagg agaugagggc 1500 aacagcagag gacuucaaca agguuaugca agucguccga gagcaaauca cucgagcuuu 1560 gcccuccaaa cccaacucuu uggaucaguu caagagcaaa uugcguagcc ugaguuacuc 1620 ugagauucua cgacugcgcc agucugagag gaugagucag gaugacuucc aguccccgcc 1680 aauuguggag cugagggaga agauccagcc cgagauccuu gagcugauca agcagcagcg 1740 ccugaaccgg cucugugagg gcagcagcuu ccgaaagauu gggaaccgcc gaaggcaaga 1800 acgguucugg uacugccggu uggcacugaa ccacaagguc cuucacuaug gugacuugga 1860 ugacaaccca caaggggagg ugacauuuga aucccugcag gagaaaauuc cuguugcaga 1920 cauuaaggcc auugucacug ggaaagauug uccccacaug aaagagaaaa gugcucugaa 1980 acagaacaag gagguguugg aauuggccuu cuccauccug uaugacccug augagaccuu 2040 aaacuucauc gcaccuaaua aauaugagua cugcaucugg auugauggcc ucagugcccu 2100 ucuggggaag gacaugucca gugagcugac caagagugac cuggacaccc ugcugagcau 2160 ggagaugaag cugcggcucc uggaccugga gaacauccag auucccgaag ccccaccccc 2220 cauccccaag gagcccagca gcuaugacuu ugucuaucac uauggcugag ccuggagcca 2280 gaaacgacgg uacccaggag aagggauuuu gggcccagga gaaacacuua cauucuggug 2340 ccuugucuuu ugcuuguaca gaaucuguag ugauuuuggu ggccaguaaa ugccagccau 2400 uucucaaacc caccucggac cacccagagu uuccucuugg ucccugucua cuaagaguca 2460 ugaaggcagg gugcucugcc cacuccauca ccaugaagcc ugggauuggg ccacgaggaa 2520 caaacagcag augcccuugc cuuccagucc aagaaacugc uucuugaaau ggauuuaaca 2580 acagccacuc accuuuuccu ccugagccug cucucugauc agcuggaucc ccacgugagc 2640 aacagcuggc ccaggaaagg cugccugcag aggacaggug uguugggcgu guugagagcc 2700 uugaagugac uaccuguauc uuagaucuga guacaagccu gaggcuuuug cuuuugucuu 2760 uuuugaugag ggcucacucc agcuucauau ggugccaaga cguugcugcu ucugagguug 2820 gcucuaacau cucuggucuu uagagccacc agaucucucu ggcccauaca gauaucagag 2880 cagacggaaa uuucucccug caagcgcuca gucucauccc agcaagucaa agaccuccug 2940 gccaaguccu gcccucuuaa gucuccagga acgcugcagg gaaaacccag cugaggccug 3000 ggccuagacu guggugaggu cacuagauuc uacugcucuu cccccacauu aauaccuuuu 3060 ccuuccucag agagaaaucu ccccuaaccu gaauugcagc ccccuccagu uugcuuuccu 3120 uuggccuucc agaccccagg aaguuggccu ucccuuccua gugcuauggu uucugccauu 3180 ggccaugauu ucagggagcu ggcugaggcc ggcugaggcc acaccugugc caguggggcu 3240 ucccuggugc ugcagcacuu guaaaccaca cacacagccu cucucccugg acauacguua 3300 gcacauuggc auucaguauu gguggccugg caugguaggu acuacccaau gaagagugua 3360 cuauauauuu ucauuacuau aggccauacu uauacagacg uguauauaua uuuauauaag 3420 aucuaccuau cuuaggaugg aaccuugggg aaaaauaaaa uugaggggaa guaaaaagua 3480 uguaacacuu ccaguuguga gccaagauug uaaccagaga gcagccagga gcuuccuguc 3540 aguaaccaug uuuucaauaa auacucuuuc auguacaaaa aaaaaaaaaa aaaaaa 3596 <210> 67 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> ELMO2 sense primer <400> 67 ctctagaaga ccccaggaag ttggcctt 28 <210> 68 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> ELMO2 antisense primer <400> 68 gaattccccc aaggttccat cctaaga 27 <210> 69 <211> 3599 <212> RNA <213> Homo sapiens BAP1 mRNA <400> 69 gcccguuguc uguguguggg acugaggggc cccgggggcg gugggggcuc ccgguggggg 60 cagcgguggg gagggagggc cuggacaugg cgcugagggg ccgccccgcg ggaagaugaa 120 uaagggcugg cuggagcugg agagcgaccc aggccucuuc acccugcucg uggaagauuu 180 cggugucaag ggggugcaag uggaggagau cuacgaccuu cagagcaaau gucagggccc 240 uguauaugga uuuaucuucc uguucaaaug gaucgaagag cgccgguccc ggcgaaaggu 300 cucuaccuug guggaugaua cguccgugau ugaugaugau auugugaaua acauguucuu 360 ugcccaccag cugauaccca acucuugugc aacucaugcc uugcugagcg ugcuccugaa 420 cugcagcagc guggaccugg gacccacccu gagucgcaug aaggacuuca ccaaggguuu 480 cagcccugag agcaaaggau augcgauugg caaugccccg gaguuggcca aggcccauaa 540 uagccaugcc aggcccgagc cacgccaccu cccugagaag cagaauggcc uuagugcagu 600 gcggaccaug gaggcguucc acuuugucag cuaugugccu aucacaggcc ggcucuuuga 660 gcuggauggg cugaaggucu accccauuga ccaugggccc uggggggagg acgaggagug 720 gacagacaag gcccggcggg ucaucaugga gcguaucggc cucgccacug caggggagcc 780 cuaccacgac auccgcuuca accugauggc aguggugccc gaccgcagga ucaaguauga 840 ggccaggcug caugugcuga aggugaaccg ucagacagua cuagaggcuc ugcagcagcu 900 gauaagagua acacagccag agcugauuca gacccacaag ucucaagagu cacagcugcc 960 ugaggagucc aagucagcca gcaacaaguc cccgcuggug cuggaagcaa acagggcccc 1020 ugcagccucu gagggcaacc acacagaugg ugcagaggag gcggcugguu caugcgcaca 1080 agccccaucc cacagcccuc ccaacaaacc caagcuagug gugaagccuc caggcagcag 1140 ccucaauggg guucacccca accccacucc cauuguccag cggcugccgg ccuuucuaga 1200 caaucacaau uaugccaagu cccccaugca ggaggaagaa gaccuggcgg cagguguggg 1260 ccgcagccga guuccagucc gcccacccca gcaguacuca gaugaugagg augacuauga 1320 ggaugacgag gaggaugacg ugcagaacac caacucugcc cuuagguaua aggggaaggg 1380 aacagggaag ccaggggcau ugagcgguuc ugcugauggg caacugucag ugcugcagcc 1440 caacaccauc aacgucuugg cugagaagcu caaagagucc cagaaggacc ucucaauucc 1500 ucuguccauc aagacuagca gcggggcugg gaguccggcu guggcagugc ccacacacuc 1560 gcagcccuca cccaccccca gcaaugagag uacagacacg gccucugaga ucggcagugc 1620 uuucaacucg ccacugcgcu cgccuauccg cucagccaac ccgacgcggc ccuccagccc 1680 ugucaccucc cacaucucca aggugcuuuu uggagaggau gacagccugc ugcguguuga 1740 cugcauacgc uacaaccgug cuguccguga ucuggguccu gucaucagca caggccugcu 1800 gcaccuggcu gaggaugggg ugcugagucc ccuggcgcug acagagggug ggaaggguuc 1860 cucgcccucc aucagaccaa uccaaggcag ccaggggucc agcagcccag uggagaagga 1920 ggucguggaa gccacggaca gcagagagaa gacggggaug gugaggccug gcgagcccuu 1980 gaguggggag aaauacucac ccaaggagcu gcuggcacug cugaagugug uggaggcuga 2040 gauugcaaac uaugaggcgu gccucaagga ggagguagag aagaggaaga aguucaagau 2100 ugaugaccag agaaggaccc acaacuacga ugaguucauc ugcaccuuua ucuccaugcu 2160 ggcucaggaa ggcaugcugg ccaaccuagu ggagcagaac aucuccgugc ggcggcgcca 2220 aggggucagc aucggccggc uccacaagca gcggaagccu gaccggcgga aacgcucucg 2280 ccccuacaag gccaagcgcc agugaggacu gcuggcccug acucugcagc ccacucuugc 2340 cguguggccc ucaccagggu ccuucccugc cccacuuccc cuuuucccag uauuacugaa 2400 uagucccagc uggagagucc aggcccuggg aaugggagga accaggccac auuccuucca 2460 ucgugcccug aggccugaca cggcagauca gccccauagu gcucaggagg cagcaucugg 2520 aguuggggca cagcgaggua cugcagcuuc cuccacagcc ggcuguggag cagcaggacc 2580 uggcccuucu gccugggcag cagaauauau auuuuaccua ucagagacau cuauuuuucu 2640 gggcuccaac ccaacaugcc accauguuga cauaaguucc uaccugacua ugcuuucucu 2700 ccuaggagcu guccuggugg gcccaggucc uuguaucaug ccacgguccc aacuacaggg 2760 uccuagcugg gggccugggu gggcccuggg cucugggccc ugcugcucua gccccagcca 2820 ccagccuguc ccuguuguaa ggaagccagg ucuucucucu ucauuccucu uaggagagug 2880 ccaaacucag ggacccagca cugggcuggg uugggaguag ggugucccag ugggguuggg 2940 gugagcaggc ugcugggauc ccauggccug agcagagcau gugggaacug uucaguggcc 3000 ugugaacugu cuuccuuguu cuagccaggc uguucaagac ugcucuccau agcaagguuc 3060 uagggcucuu cgccuucagu guuguggccc uagcuauggg ccuaaauugg gcucuagguc 3120 ucugucccug gcgcuugagg cucagaagag ccucugucca gccccucagu auuaccaugu 3180 cucccucuca gggguagcag agacaggguu gcuuauagga agcuggcacc acucagcucu 3240 uccugcuacu ccaguuuccu cagccucugc aaggcacuca ggguggggga cagcaggauc 3300 aagacaaccc guuggagccc cuguguucca gaggaccuga ugccaagggg uaaugggccc 3360 agcagugccu cuggagccca ggccccaaca cagccccaug gccucugcca gauggcuuug 3420 aaaaagguga uccaagcagg ccccuuuauc uguacauagu gacugagugg ggggugcugg 3480 caaguguggc agcugccucu gggcugagca cagcuugacc ccucuagccc cuguaaauac 3540 uggaucaaug aaugaauaaa acucuccuaa gaaucuccug agaaaaaaaa aaaaaaaaa 3599 <210> 70 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> BAP1 sense primer <400> 70 ctctagacct gactctgcag cccactctt 29 <210> 71 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> BAP1 antisense primer <400> 71 gaattcagga agagctgagt ggtgccag 28 <210> 72 <211> 4611 <212> RNA <213> Homo sapiens WDR37 mRNA <400> 72 gggucgcguc agugcggaga cagcgguguc cugcgcgucu ucgggucagu acggggggcc 60 gcgucguagg gacucacugg cggugcgcga ccugggcugg cuccgggggu ggcgggcgca 120 gcugcuguga cagcuuauug caggagugac caggacacua ccuccuagaa guaaugccca 180 cagaaagcgc aaguuguucg acugcucgcc aaacaaaaca gaagcgcaaa ucccauagcc 240 uuucuauacg aagaacuaac agcucggagc aggagaggac gggacugcca agagacaugu 300 uagaaggaca agauucuaaa cugccuuccu cgguucgcag uacacuucug gaacuguuug 360 gucaaauaga aagagaauuu gaaaaccuuu auaucgaaaa cuuagaauua cguagagaaa 420 ucgacacucu uaaugaacgu uuagcugcug aaggacaagc gauugaugga gcagagcuga 480 guaagggcca acucaaaaca aaagccaguc acagcaccag ccagcucucc cagaaacuga 540 agaccacuua caaggcuucc accagcaaga uugucuccag cuuuaagacc acgacaucga 600 gagcugccug ccagcucgug aaggaguaca ucggccaccg ggacggcauc ugggauguca 660 gcguggccaa gacacagcca guggugcucg ggacugcauc agccgaucac acggcuuugc 720 uguggagcau agagacaggg aagugccuag ucaaguacgc aggccacgug ggcucaguaa 780 auucuaucaa auuucauccc ucagagcagu uggcucucac ugcuucugga gaucagacug 840 cucauaucug gagauacgcg gugcagcugc cgacacccca gccuguugcu gacacuagua 900 uaucugggga agaugaagua gagugcucug acaaggacga gcccgaccuc gauggggaug 960 uguccagcga cugccccacc auccgcgucc cacugacauc ccucaagagc caccagggcg 1020 uggucaucgc cuccgacugg cugguugggg ggaagcaggc ugugacugcc uccugggacc 1080 ggacggcaaa ccuguacgac guggagacgu ccgagcucgu ucacucucug acagggcacg 1140 accaggaguu gacgcacugc ugcacacacc ccacccagcg gcucguggug accuccuccc 1200 gugacacgac uuuccgccuc ugggauuuca gggaccccuc cauccacucg gugaauguuu 1260 uccagggaca cacggacacu gugacuucug ccguguucac cgugggagac aacgugguuu 1320 caggcagcga ugaccgcacg gugaaagucu gggacuugaa aaauaugaga ucccccauug 1380 caacuauucg cacggacucu gccauuaaca ggaucaaugu augugucggc caaaaaauca 1440 uagcccuccc ccaugacaac cgacaaguga gacuguuuga uaugucagga gugcgccugg 1500 cgcggcuucc ccggagcagc cgacagggcc acaggagaau gguaugcugc ucggcaugga 1560 gugaagacca ccccgugugc aaucuguuca ccuguggguu ugaccggcaa gccauugguu 1620 ggaacaucaa caucccugca uugcuacaag aaaaauaagg acaccggcag cccuuaguuu 1680 cacuguuugc cagcacagac cuuugauggg ugcaggcuuu ucugcguauu aaucagccau 1740 uuuugugaga guuugacccu ggaaagggug cuuuguauau guucuuuuca cauagugccc 1800 agcuugcaug aaauguacag agaaaugugu ggucguauuu uuuacuuuug ucuuguauau 1860 guauguauau uggguccucu gggcaguaga ggcaaagcuc accucccaug uagcacauga 1920 aaugcuugug aguuguugac auuggacagg ugaacaguag ggcauuacau uugugugaau 1980 uaaaugugaa cuucuguauu acguugcggc gucggcaguc cugcguuccc uggaguaacu 2040 guacguaucu gccuuugcug ggaagacugu ggggcugccu guguuggcug gcgaccagca 2100 ggauugcucc aggauuuugu guuuaccucg cgugaaguuc agcacgugcu gucguguagu 2160 cagcuucuac ucuaauuucu guuacaguuc ugcaaaggua accuggaguu uagaaguuaa 2220 aaaaaagcau gggauguugg auuugcacca uuuggaguuu cuuuaggaaa gaaaaguuuu 2280 cugcuuuuuu auagaaaauc auuucagucu cccgaggucu caugcuagca aauuuugaaa 2340 uaggauucua aucacugauu ucaaauauua agcaaaaugu aaagcacuuu aauuuauagc 2400 uaugguuaua aacagguuuu agauguuuca aaugacuugu ccacugaaug ucacuugacc 2460 uugauaagag gccgccugca cacagagccc aguuaauucu ccgcaccucg guugugugcu 2520 uccgaauggg cucacucccg uggugguguu ugagagccaa caacacuacc ucagagacgg 2580 gucuuuggga aacuuugggu cucacuguug ccuggcugga gcacuuuggu uuauagcugg 2640 aauacugagu ucaguucaga aggcaggaaa gacagucaca ccgacguguc cugaaggugu 2700 aggcucucca cuuaggcgca caagcugacg gcugcagcca gcaggccccg gugacgagac 2760 acuuccaggu cuuguggugg ggacgccucu cagugccagu cccgccacug cugagugagc 2820 cugguguucu ugccuucuug gaaauuacug cucaccuggu aucuguacgu uaauguuucu 2880 ugcugaguua caguuuugau aaagaggcuc ucauuuccug ugucuuguau auucaguccu 2940 uucaauacgu ccaccuggag gcucaccacu uggagagaca caggaaggua auauuuacag 3000 cugucaugug acauccccag gucuuugugu uuugcccugu uuuacgguga gguaggaggg 3060 aacccaucug gggaccggua ggugcaggug caguaggacg ugggacuuuu ggacccgucc 3120 uuuggugcag cucgccaggg augagaggca ccucccuacu ugggucuuca ggagcugguc 3180 caaggagcuu cgaaucuaag ucaucuagaa ugacccugaa augacugaca gccccgggcc 3240 caagaaaaac ccauaaccac cucagaugga ucugacgugg cuaagggaca aacagcaaau 3300 auuucaguca uuuugauuuu acaaauaaaa aauguguugu guuuuugucc gacauuauuu 3360 ccugacugca cuguucugag aauggagucc accugguccc ucugguugau uagaaucuca 3420 gguuucagcu ccugcugucc ugagcgaacu ugccugaugc agggcugugc uguguccaga 3480 uguugcuggg gccucacuuu uucucuuggc uggaggucca auugccagag ccucccacac 3540 ugcacauaca aaggucugag cccagggcag cuucuggggc cacugcacag gccaccugcu 3600 uggguuccuc ggaguuuaau uugaaagucu gggugucuua ggaugauggu uaggaacauu 3660 gaaaaauggc ugcaaauagc caaaucaaac uuaagaacca gaucucugcc agauuaaaca 3720 uuuuugaagc uuuuaaaagu caauauuccu aguggccacu gaguuccagg cacacuggug 3780 cccuuuacug ccacagcugc ucaccuuguc uggcaaacug gagggaccuc agaaacugga 3840 cuccugcaug uccuuggggg cgcagcccug uggugcucag gcagagcucu caggagccgg 3900 ggcaccuugc uguucgcugc ugugucgucu ucuaauguga gcucauccac ugcugcugca 3960 gcguggugau caggagucac agacaagauc ggggauggug ugugugugug ugugugugug 4020 ugugugcacg uguguguggc uaaauuaagu cauacuguca accacacgug aucucgucug 4080 aaacaguguu uggaaguggg aacaguuuug uccuguaugc ugaugugucc agaauuucau 4140 uuaaugauag acggaaaaug ugugguuacu gaaaacugua uaugauacag aauuucauaa 4200 gagccaugcu guugggcaaa gcaacucuuu uucaaccacu gcucaucagu uucuguagag 4260 acaaaaacuc uguacauauu uuggaaucug aagaauccua uguaaaucau uuguuacuua 4320 agucugugaa aaacauauuu cuuuggagga aaauguaugc auuuauaagu guuccaugga 4380 aucaguuuuu auuguaucga uauaauuguc ucuaaguguu gacugucuuc auugcaauau 4440 gaaauucauu aaaaugucca uguuccauaa uuacuauuau aaaagcuuug uguuauuggg 4500 agucuuauua uuuuuauagu uuuucauguu uugcuuuggu gaaacauugu gaaugacuau 4560 auaaacauca cgaugagcua gaaauugaac aaauauaaua cuggucacuc g 4611 <210> 73 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> WDR37 sense primer <400> 73 tctagagtct gggtgtctta ggatgatgg 29 <210> 74 <211> 31 <212> DNA <213> Artificial Sequence <220> <223> WDR37 antisense primer <400> 74 tctagagttc aatttctagc tcatcgtgat g 31 <210> 75 <211> 3400 <212> RNA <213> Homo sapiens KLHL20 mRNA <400> 75 augggccggu gaagggaaag gaaauagcuc uacgagcggc auccugccug cguuagcggc 60 gguggaggag gaggcagaga ggaguggagg gcggaguaga cggaggaggc ugcugcagag 120 aagaaagugu cagagccggu guaccaacau ucgaccagga gagacuggaa uggauguaac 180 aagccgcugc acccuuggag accccaacaa acugccagaa gggguucccc aaccugcccg 240 caugcccuau aucucagaca agcacccucg acaaaccuug gaagugauua accuucugag 300 aaagcaccgg gagcuaugug auguggugcu aguugugggc gccaagaaga uauaugccca 360 ucgagucauu uugucagccu guagucccua cuuccgagcu auguuuacag gagaauuggc 420 agagagccgu cagacagaag uagugauccg agacauugac gagagggcua uggaauuacu 480 gauugacuuu gcguauaccu cccagauaac aguagaagag ggcaauguuc agacucuucu 540 gccagcugcu ugccuccucc agcuggcaga aauacaggaa gccugcugug aauucuuaaa 600 gagacaauua gauccuucua acugccuggg cauucgggcu uuugcugaca cacauucaug 660 ucgugaguug cuaaggauag cagacaaguu cacccaacau aacuuucaag agguaaugga 720 gagugaagag uucauguugc uuccagccaa ucaacucauu gauauaauau ccagugauga 780 gcuaaacguu cgcagugaag aacaaguguu caaugcagug auggccuggg ucaaauacag 840 uauucaggaa agacguccuc aauuacccca ggugcugcag cauguucguu ugccuuugcu 900 uagucccaag uuccuggucg gcacaguagg cucugauccc cucaucaaaa gugaugaaga 960 augcagagac uugguagaug aggcuaaaaa cuaccuccua uugccgcaag aacgaccacu 1020 aaugcaagga ccaaggacga gaccacggaa accuauccga uguggggaag uacucuuugc 1080 aguugguggu uggugcagug gagaugccau uuccaguguu gaacgauaug auccacagac 1140 caaugaaugg agaauggugg cuucaaugag caaaaggaga ugcggaguug gggucagugu 1200 ucuugaugau cuguuauaug caguaggagg ccaugaugga uccucuuauc ucaauagugu 1260 ugaaagguau gaccccaaaa caaaccagug gagcagugau guggccccua caagcaccug 1320 caggacaagu guugguguag caguacuugg aggcuuucuu uaugcugugg guggccagga 1380 uggugugucu ugccucaaca uuguugagag guaugauccg aaggagaaca aguggacucg 1440 gguagcuucu augaguacca gaagacuagg uguggcugug gcuguguuag gaggguucuu 1500 auaugcugua gguggcucug acgggacauc uccucucaac acaguggaac guuacaaucc 1560 ucaggaaaac agauggcaca cuauagcccc uauggggacc cggaggaaac accuaggcug 1620 ugcaguauau caggacauga ucuaugcugu aggagguaga gaugacacua cagagcugag 1680 cagugcugag agauacaacc ccagaaccaa ccaguggucu ccaguggugg ccaugacauc 1740 acgccguagu ggaguuggcc uggcaguggu caauggacag cucauggcag uaggagguuu 1800 ugauggcaca acauacuuga agaccauaga aguuuuugau ccugaugcca auacauggag 1860 guuauauggc gggaugaauu accgucggcu aggggguggc guaggaguua uuaaaaugac 1920 acauugugaa ucccauauuu ggugaacaca gagaagacag ucuuguauau auuccucugu 1980 auucugggga gcuuugaccu uggagcuuug uacagcuuga gaaaacauua gaacaaauuu 2040 uauuauuugc cggugccuca acaaauggaa auacaaucca augaaaguac uucaccugca 2100 agaugcacaa uaauuuucaa cucugugcag aagaauauuu auuuuugguu uuaauuuauc 2160 augguuuuuu guuguuuucg uuuugaacuu auccuuccuc ccacaaaaaa agaaaagaag 2220 aaaaaauucc aagcagcaaa acuuacuuug uuuguaaggu auucauuuag guuugaaaau 2280 acuauuuaau aagggcagaa gggcuauaua ugauuuggcu auuauuucua gacacuccau 2340 ccacaugauu ccacuaacaa ggauuaccag gaauaaaggu agguaugcaa aauguauuag 2400 cuacccauua uucucgcacu aaccaccaga agacuugaaa aucuuaaaaa aaaaaaacaa 2460 aaaaacaaga aaaggcaaca ucucauuuua aauaguacaa uucagaaggg auacuaaauc 2520 aauaaaaacc aggacucaga cacuacaguu uucaucagug uaauuuuaug ucuuguuucu 2580 uucuauauga acuuguuuau uuagucuuuu uuucauaaua uucucauaga guuucugcua 2640 gaucuagagc ucugcucagu gccucuuaua aaaacaauau auaagucuca uaugcugucu 2700 ugagaaucca agagaaucac augcagcucu gcaacaguuu uugcucaaaa auguuacuga 2760 cuaaagcaac uguccuuccu guuuccuuau ugcuaccaag gaccagcagg gagaaauguc 2820 uucucccagc agugaauucu guuaugcaau uuauucuuga ugcucaggcc ugguuaagcu 2880 gaggcuuacc uuuaauaaca gccuccaagg gaaugaauua uucaguuaau guaauagcac 2940 acauuaaaga guguaaaauc aauugaggca uuuuauuaau aucuugguuc uuuuauacaa 3000 acaauguauc auuaucuuca uucuauaagg ucauuguuac uacuucucuc aaauugcacu 3060 uucugguaaa aaguuaaaaa uuuuuaagga aaaaauaauu ucaaugguua aaguuuuuuu 3120 uccucauugu auucauagac aaauuuuuuu caauauauuu ugcaagaaaa aucuuugaau 3180 aagacuaacu gcauuuaaaa ggaauaaccu ccuucucccu uuccccaacu acaaaaaugu 3240 uuggcaacuu uguguuuaca uuuaaagauc auuugcaccu uuuucaagga aaaaaaguau 3300 ugaguaaaca auuguuuaca uauaucauuu augcuuuuuu cuagcaugua uaacuuuuuu 3360 aaauaaaggu aguauuuacc auuaaaaaaa uuuuagcuaa 3400 <210> 76 <211> 31 <212> DNA <213> Artificial Sequence <220> <223> KLHL20 sense primer <400> 76 tctagaagaa aaatctttga ataagactaa c 31 <210> 77 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> KLHL20 antisense primer <400> 77 tctagaagct aaaatttttt taatggtaa 29 <210> 78 <211> 10051 <212> RNA <213> Homo sapiens PTPRD mRNA <400> 78 gcaguuuauu cugggccugu guggaauaga aagacaauuc uuuauuugac ugguaggaaa 60 gaaaagagaa gacuccucag gguuuccaag gaggacugga gcggcugcuu uagugaagaa 120 gugaagaauc aaggaaguaa aggaccuguc ucaaagucac acaacuagga aaucuugagg 180 agucugauac uuuucauccu ccagcuauaa aaucaucuug uagggggaag cuguacauug 240 guggaaaaua aaaggcaagc aguuccuggc ccgugauauu ccaguuagag gugucugacu 300 gacagcaugu uggcaccgag ggaagcgaug ggcuucggcu ccagguuucu guccugagac 360 cacagcagaa ugugccuugg acaaaggcuu ggacuccuug aucuucuggu aacuucaaaa 420 aucuggucuc agggugguga acuuuuucuu cgaauuccua accugugagc uuacgaugau 480 ggugccuguc cauuaacuga acaacugugg uuuugaacuu cuugaggcca gggacugugu 540 cugaaaacac cgacgguggc uaacucaagg gagacgucug gugaacaccc gugggaucua 600 aagaacaagc ucugaaagug uuccagcuga aauuucagau cggacagacu cgcugcggcu 660 ccggaggcag ugauuccaag cugcucgcgc acgcugcugc caagcugcag gauggugcac 720 guagccaggc ugcugcugcu gcuccucacu uucuuccucc gcacggaugc ugagacaccu 780 ccaagguuua cacgaacacc cguugaucag acaggggucu cuggcggagu ugccucuuuc 840 aucugccaag cuacgggaga cccaagaccu aaaauugucu ggaacaaaaa aggaaagaaa 900 gucagcaauc agagauuuga gguaauagag uuugacgaug ggucuggauc aguucucaga 960 auacaacccu uacggacucc gagggaugag gccauuuaug aauguguggc cucaaauaau 1020 gugggagaaa uaaguguauc caccagacuc acaguuuugc gggaagauca aauucccagg 1080 ggcuucccua ccauugacau gggcccacag uugaaggugg uugagcguac ucgcacggcc 1140 accaugcuuu gugcagccag ugguaauccg gauccagaaa ucacuugguu uaaagauuuc 1200 uuaccugugg acacaagcaa caacaauggu cguauuaagc aguuacgauc agaaucuauu 1260 ggugguacac caauaagagg agcccuucag auugagcaga gugaagaguc ugaccaagga 1320 aaauaugagu guguugccac caacagcgcg ggcacucgcu auuccgcucc ugccaauuua 1380 uaugucagag agcugcgaga aguucgccgu gucccaccaa gauucucuau cccacccacu 1440 aaucaugaaa ucaugccagg cggaagcguu aauaucaccu guguggccgu ggggucacca 1500 augccuuaug uaaaguggau guugggggca gaagaucuga caccugaaga ugauaugcca 1560 auaggaagaa augugcuaga acugaaugau guaagacagu cagcaaauua caccuguguu 1620 gcuaugucaa cacugggugu cauugaagca auagcacaga ucacugucaa agccuuaccc 1680 aaaccuccag gaacuccugu agugaccgag agcacagcua caagcaucac acugacgugg 1740 gacucuggga acccugagcc uguuucuuau uacauaauuc agcauaaacc uaaaaacucu 1800 gaggaacuuu acaaagaaau ugauggggug gcgaccacac gcuacagugu cgcuggacua 1860 agucccuacu cggauuauga auucaggguu guugcuguca auaacauugg gcgggggccu 1920 cccagcgaac cugugcuaac acaaaccuca gagcaagcac cauccagugc cccgagggau 1980 guccaggcac gaauguugag uucgaccacc auuuugguac aguggaagga accugaagag 2040 ccaaauggac agauccaagg auauagaguu uauuauacaa uggaucccac ucaacauguc 2100 aacaacugga ugaaacacaa uguagcugac agccaaauca cuacuauugg caacuuagug 2160 ccccagaaaa cauauucugu caaaguccug gcuuuuaccu caauuggaga ugguccccuu 2220 ucaagugaca uacaagucau cacucagaca ggaguaccag ggcagccacu aaacuucaaa 2280 gcagaaccug agucugaaac aaguauuuug cucucuugga caccuccacg uucagauacc 2340 auugccaacu augaacuggu cuacaaagau ggggagcaug gagaggagca acgaauuacc 2400 auugagccag ggacaucaua uaggcugcaa ggacugaaac caaacagcuu auacuauuuc 2460 cgucuggcug cacgcucccc ucaaggccug ggugcuucua cugcagaaau aucagcuaga 2520 accaugcagu caaagccguc agcuccuccu caagacauua guugcaccag cccaaguucc 2580 acuaguauuu ugguaaguug gcaaccucca ccaguggaaa aacagaaugg cauuaucacu 2640 gaauacucca ucaaguacac ugcaguggau ggggaagaug acaagccuca cgagauuuug 2700 ggaauuccuu cggacacuac caaauaccuu uuggaacagc uggaaaaaug gacugaauac 2760 cggaucacug ugacagccca uacagauguc ggcccuggcc cugagagcuu guccguguug 2820 auucgaacca augaagaugu uccuaguggu ccuccucgca aagucgaggu agaggcuguc 2880 aacucaacau cuguuaaagu cucauggcgc ucacccgugc ccaauaaaca gcauggccag 2940 auaagaggau aucaggugca uuaugugagg auggaaaaug gugagcccaa gggccagccc 3000 augcugaaag augucaugcu ggcugaugca caggacauga ucauuucugg gcuccagccu 3060 gaaacuuccu acucccucac cgucacagcc uacacaacca aaggagaugg ugcucgcagc 3120 aagcccaaac ugguguccac cacuggggca guuccaggga aaccucggcu ugugauuaac 3180 cacacucaga ugaauacugc ucuuauucag uggcacccuc cgguggacac auuuggaccu 3240 cuucagggcu accgucuaaa auuuggccgc aaggauaugg agccacuuac uacucuugag 3300 uucucugaaa aagaagauca cuuuacagcu acagacaucc acaagggagc aucauacguc 3360 uucaggcucu cagccagaaa caaagugggc uuuggggagg agauggugaa ggagauuucc 3420 auuccagaag aaguaccaac uggauucccu caaaaccuuc acucagaagg caccacuuca 3480 accuccgucc aguuaucuug gcaaccaccu guccuggcag agagaaaugg cauuaucacc 3540 aaguauaccc uucuuuauag ggauaucaac aucccccuuc ucccgaugga gcagcuuauu 3600 guuccagcug acaccacuau gacacucacu ggcuuaaaac cagauaccac auacgaugua 3660 aaaguacgug cucauacgag caaagggccc gggccauaua gucccagugu ccaguucagg 3720 acacugccug uggaucaagu guuugcaaaa aauuuucaug ucaaagcagu aaugaagacu 3780 uccguguugc ugucuuggga gauuccagag aauuauaacu ccgccaugcc uuucaaaauu 3840 cuuuaugaug augggaaaau gguagaagaa guggauggcc gagccacaca gaaguuaauu 3900 gucaaccuga agccugagaa aucauauuca uuugugcuga caaaucgugg aaacagugcu 3960 ggugggcugc agcacagggu cacggcaaag acugcaccag auguauuacg uaccaagccu 4020 gccuucauug ggaagaccaa cuuggauggc augauuacug ugcaacugcc ugaaguaccu 4080 gcaaaugaga auauaaaagg uuacuacaua auaauugugc cuuugaagaa aucucgcggg 4140 aaauuuauca agccauggga gaguccagau gaaauggaau uagaugagcu gcuuaaggag 4200 auaucuagga agcgcagaag cauccguuau gggagagaag uugaauuaaa gccauauauu 4260 gccgcucacu uugauguccu ucccacugag uucacccugg gggaugacaa gcauuauggu 4320 ggauuuacaa acaagcaacu ccaaaguggu caagaauaug ucuucuuugu guuagcagua 4380 auggaacaug cagagucuaa gauguaugca accagcccuu acuccgaccc cgugguguca 4440 auggaucugg auccgcagcc aaucacggau gaagaagaag gcuugaucug gguuguaggu 4500 ccuguccuug caguggucuu uaucaucugc auugucauug cuauucuucu uuauaaaagg 4560 aagagggcag aguccgacuc uagaaaaagc agcauaccga acaauaagga gaucccuuca 4620 caccacccaa cagacccugu agaacugagg cgccuuaacu uucaaacacc ggguauggcu 4680 agccauccuc caauacccau cuuggaacuu gcagaccaca uugaaagauu gaaagcaaau 4740 gacaacuuga aguuuuccca ggaauaugag ucaauugacc cuggccagca guucacuugg 4800 gaacauucaa acuuggaagu aaacaaacca aagaauagau acgcgaaugu aaucgcauau 4860 gaucauuccc ggguucuccu aucagcuaua gaagggaucc caggaaguga cuaugugaau 4920 gccaacuaca uagaugggua uaggaagcaa aaugccuaua uugcaacaca gggaucucuc 4980 cccgaaacau uuggggacuu uuggagaaug auaugggaac aacggagugc cacaguuguc 5040 augaugacaa aacuagaaga aagaucaagg gugaagugug accaguauug gccuagcaga 5100 ggcacagaaa cccacggacu cguucaagua acgcugcuug auacugugga gcuggccaca 5160 uauuguguuc gaacauuugc acuuuacaag aaugguucaa gugagaagag agaagugaga 5220 caauuccagu ucaccgccug gccugaucau gguguuccag aacacccuac accuuuucua 5280 gcuuucuuac guagagucaa aaccuguaac ccucccgaug cugguccgau gguugugcac 5340 ugcagugcgg gaguuggccg gacugguugc uucaucguca uagaugccau guuagaaaga 5400 auaaagcaug aaaaaacugu agauauuuau ggccauguaa cuuuaaugag agcccagagg 5460 aacuauaugg uucaaacaga agaccaauac aucuuuaucc augaugcacu guuagaagca 5520 gugacuugug gaaauaccga agugccagcu agaaacuugu augccuacau ucagaagcug 5580 acacaaauag aaacgggaga gaaugucaca ggaauggagc ucgaauuuaa gcgucuagcc 5640 agcucaaaag cucacaccuc aagguuuauc agugccaauc uuccauguaa uaaauucaaa 5700 aaucgccuug uuaauauuau gccauaugaa uccacaaggg uaugccugca gccuauccgu 5760 ggaguagaag gaucugauua caucaaugcc aguuuuauug auggauacag acaacagaaa 5820 gccuacaucg cuacccaggg gcccuuggca gagaccacug aagacuucug gcggaugcuc 5880 ugggaacaca auuccaccau aguugugaug cucaccaagc ugcgugaaau gggcagagag 5940 aaaugucacc aauacuggcc agcagaacgg ucugcaagau accaguacuu uguuguagau 6000 cccauggcug aguacaacau gccacaguau auccuaaggg aauucaaggu cacagaugcc 6060 agggacggcc agucccgaac aguaaggcag uuccaguuca cugacuggcc agagcaagga 6120 gugccaaagu ccggagaagg auuuauugac uucaucggcc aaguccauaa aacaaaagaa 6180 caguuuggcc aagauggacc cauuucaguc cauugcagcg cgggcguugg aagaacugga 6240 gucuucauaa cgcuaagcau uguuuuggaa agaaugagau augaaggagu uguagauauc 6300 uuccagacug ucaaaauguu aagaacacaa cgaccagcua ugguacagac agaggaucaa 6360 uaucaguuuu ccuaucgugc cgcacuagag uaccugggca gcuuugacca cuaugcaacg 6420 uagaaacccc ugacccauuc uggauuuuua cuacaggccc uucaauaucc auggagucuc 6480 uucugagcca uacagggcac uugagaaguc cuucuuaacu ucuagcuaac aacuacuuag 6540 ugggacuauu acacacaaaa caaauuaaaa acaaauuauu ccagguggac caagaauuca 6600 caguuuaaua aucuaaccug aaaaauaaug aagagaaucc ugacugauga ggcaucugaa 6660 ggauuuuauu uacagauacc ucaaggauuc aaaaucaagg gaagaagaaa ucacagcaaa 6720 gaaccaccau cuuauucagg auugcuugaa aggugcaagg agaaauugcc agaaugcugc 6780 aguucagcac aagauaugug cugguguggc uucucacaau gaaacggacu cugcuuugac 6840 aucgccccuu cccaccauac ugcucauaau aacauuuuag ggggaaaggg gagggaaugu 6900 uuaaaaagaa aguccuugau uuaguuuuuu aguauuguaa agauacugcu gaccugugcu 6960 ucauuucuaa cuguguaaac uuuuuuuuaa caaaauguau cauucgauaa agugaauuuu 7020 aaaaaaguua cauaacucuu cauuuuuuua uuuccaaauc guagguuuuu uuaagcugua 7080 gaacuguuau cuguaauauc uugcuguaga aauaucaaau cauuugaaaa uuugcacauu 7140 uuugugcaaa acucaaucua caguaucagu cucgucagau auauuaauuu uacacuucag 7200 uuuugauugg ugagaaagua cccauucucu ucaaauaauc aaagauaauu auuauuuugu 7260 uuuguuuuug gaaucaacag ggaggcgcaa aguauaaagu ugcugcuaac auauauacau 7320 auauauccau auuuuauaag ggugucuaug uauauauaga cagugugucc acacaaaaaa 7380 uagauacagu uaucagucag ucaguucuuc caugauuuag uuuuuuuaaa cguagaaaag 7440 cuauuguaaa caucucuuuc cauuuauucu uaauuuuuug acauauuggu auuucuuuaa 7500 agggaaauga ggaaugcaca ucagugauug auugucaaac cucacccccu gauuuccuac 7560 cuaaucuacc cccaccuaac caaucaauca cauccacaaa uuguuuuguu uguuuguuag 7620 ucaggcuucc aacaaaguuc aauauuucua acacucuagu gcaauaaaaa uuauuauuaa 7680 auagcuaaga ggugugcaug ugggaaaggu cagugcauau cccuuuagga ggggagaaug 7740 uuguaauaua ucagcuaucg aguuguuuaa aaaaagugua uucaaucgua uauugucuau 7800 aguaugugcu augaaauuug cauuuaugau auguaacagg ggcaaagcca aauucauguu 7860 acucuguuca gucagaaaca uuuuguggca uacagcauuc cugggaagug cuguacuuug 7920 uuucguuuug guuuuaguuu ugcauuuaga gugccuuaua auugaugccu auuuuaauag 7980 cauuucuuuu uagcuuuugg uucguauuuc cauucacugu ucguaucugu uacuuucuau 8040 uaaagcauua ucuguuuacc acauguacaa aaacucuuug aauaauaugc auuccuaguu 8100 uucagccaag ucggggaugu uagugauugu accagcccaa agcacuugga uaaucagggc 8160 ccuucuuccu uuuauaauca aucaucaaca ucagaaaaag cuacuuguuu uauuuauauu 8220 cccuuccaaa uccgcucugg aacaugcagu aacugcacca aacuuauuuu aguaacaaau 8280 aucauuggca acuuuggaau auauuugaua uuccauuagg auuuuucuaa aaggggaaau 8340 aaacuauaua uauauaugua ucuuaccccc aauucuucca acagaauuuc uauaggaagc 8400 cauggaugau ggcauaaguu ugccacauau uacaugauuu uaaauaaucc ucaaaauacc 8460 caaggaacuc uuaaagaguu uugguaugag uauacuacuu ugguuuaauu uuagcuucau 8520 ggauguucug cauggaagga uuuuuguuuu ccacauuuuc ccauugcuag cagagugaaa 8580 uccaagagac caaacauuug caagcauugu auuugagcac uuuuguaaaa aacaaagaaa 8640 aagaaaaaaa agaaaauaua uauaauacua aaaaaaagua ucuagaaggc uaccucagaa 8700 ugagacucuc uaaccuacau cagaaccaga gaagaaugug cacuaugugg gucuguuauc 8760 auuauuuucu uuuaguuugu aucuuuuugg agauuuaucc aagugccaga uuacucagug 8820 cuauaauuuu cuuuuaguua aacaaagggg gucagacaga cauugcauca uccagacaug 8880 ccuuguugga cauguagaau ccgauggagc acugcacacc agaaugauug gccaaugagc 8940 agcuucucuc ccugaaacaa uaacugccca uuuggcaaag ggaaagauga caauaaucag 9000 aagaagaaaa ugaaugggau gcauaccaua gacgaacgag gcggagacua uugcgggaau 9060 cuuacuguuc aggagcuguu ccuagaacua acucccuuac ugucauugau gugcauucca 9120 cucugugcuu uucuguacaa ccauucaagu uuuaauuucc caggugaacc aucuuuaucu 9180 gccauuacca caagcuuuca aguuuccagu uauuuucauc aucauaacca guacggugcu 9240 auuauuuacc uauguacgug uaguuaugua uaauuuugua auuaguuaca augguaaaaa 9300 aaaucaaaau auauaaaaag ugauuuguac agaacuuuau uuuagcucuu uuuuaaaaau 9360 gauuugcaug guuagaaaac ggcgaggaca gccaggggag ggaagggccu cuagggaacu 9420 uugcacuuuc uauaccuuug uacuaugcac ugcccuauug auucuacacc caauaaugau 9480 auuacuugaa cccaucugua agaaacugcu ucggaaauuc auuugugugu auguaaauaa 9540 cacaacauag aaacaggaag ggaaaaaagu cugcaguaau gcacguauuu uuuuucuuuc 9600 cuguuuauuu ucgguuuugc uuuaaguccu uuuauuuuua auucccuuuu uguuuuucuu 9660 uuuggguuuu gguuccuuuu ggguuuaugg gugcccugau acuccagcag agaucagaag 9720 gcuacagauc cauucuaucc auccguuaug uggcuuugcc aucccagcuu ggagugucuu 9780 uacaaagaua auaacaguug uguucuuugc ucucguuuug gaugcauaga cugaaaaauu 9840 aaaacaaaua acuuguaaaa uggcuuguua aaaaauacaa uuaccucuaa uuaguaguac 9900 gcguaaaugu uuuacagaau gaaaggcgug cuuuuuauuu ucuuacuucg uuacauuggu 9960 ggcgaaagaa gucuguauga aaaucaguuc uuugcugaca caaguuccau uuguuacaaa 10020 Ugaauucuaa uaaaaauguc aguguuaugc a 10051 <210> 79 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> PTPRD sense primer <400> 79 ctctagacgt tatgtggctt tgccatcc 28 <210> 80 <211> 30 <212> DNA <213> Artificial Sequence <220> PTPRD antisense primer <400> 80 gaattcgtaa caaatggaac ttgtgtcagc 30 <210> 81 <211> 5307 <212> RNA <213> Homo sapiens RASSF2 mRNA <400> 81 aaugcuguga guuccuuguu aggccucucc uuccgaugaa aagagaaagg aagaauggac 60 uacagccacc aaacgucccu agucccaugu ggacaagaua aauacauuuc caaaaaugaa 120 cuucucuugc aucugaagac cuacaacuug uacuaugaag gccagaauuu acagcuccgg 180 caccgggagg aagaagacga guucauugug gaggggcucc ugaacaucuc cuggggccug 240 cgccggccca uucgccugca gaugcaggau gacaacgaac gcauucgacc cccuccaucc 300 uccuccuccu ggcacucugg cuguaaccug ggggcucagg gaaccacucu gaagccccug 360 acugugccca aaguucagau cucagaggug gaugccccgc cggaggguga ccagaugcca 420 agcuccacag acuccagggg ccugaagccc cugcaggagg acaccccaca gcugaugcgc 480 acacgcagug auguuggggu gcgucgccgu ggcaauguga ggacgccuag ugaccagcgg 540 cgaaucagac gccaccgcuu cuccaucaac ggccauuucu acaaccauaa gacauccgug 600 uucacaccag ccuauggcuc ugucaccaac guccgcauca acagcaccau gaccacccca 660 cagguccuga agcugcugcu caacaaauuu aagauugaga auucagcaga ggaguuugcc 720 uuguacgugg uccauacgag uggugagaaa cagaagcuga aggccaccga uuacccgcug 780 auugcccgaa uccuccaggg cccaugugag cagaucucca aaguguuccu aauggagaag 840 gaccaggugg aggaagucac cuacgacgug gcccaguaua uaaaguucga gaugccggua 900 cuuaaaagcu ucauucagaa gcuccaggag gaagaagauc gggaaguaaa gaagcugaug 960 cgcaaguaca ccgugcuccg gcuaaugauu cgacagaggc uggaggagau agccgagacc 1020 ccagcaacaa ucugagccau gagaacgagg ggaucugggc accccaggaa ccgccauugc 1080 ccauaagacc cccaggaagc uaggcacuuu cuuuccaugg aaacauuuag acacaaaccu 1140 ccccagcucc ggccaagcca ucauuugcua ccuggagcug gauguagaag ucagcagaca 1200 gcucccuauc ccuggacccc ugcccuccuu uuuucugcuc acaaggacuu uugauuuuag 1260 uuauaaggag gacccaaaau gugugugugu acaugugugu gcacacaugg uacgugucca 1320 ugugccuacc ugauacuuuc acauguaauu aaauuccagg caaccagcac aagagccgug 1380 agcuuggcac augugcugcu cgugagcagg aaaaucagag gagccacuga ucugaguggu 1440 auuuagguug aaggaaagau uucuccucuc aagugccagg gagcagccac acgucugucu 1500 guguuuagag agggaagagg guucuccagg uucaccauuu ggguuguuua uauguuggua 1560 gaaauucucc cuguaugccu agaaggauca gugaauguaa gagccuugga aauuaacaaa 1620 auaacagcca cauaaccuug cggcaagucu gauggaaaga aaaagauaaa ccauccgugg 1680 gguagaugca auaagcccac guauuuuuac acuggaaacg uugauuguuu uaaaugacaa 1740 agacauaugu gauguucuau guggaaaccu gugaagagug gauucugccu ccaucucugc 1800 cuccauggcu accuuuagga gacagagaag auccugugug uuucucugua cccagcugac 1860 agccugucuc uauggcgcuu ccuugagugg aaggaaaugu cucaagaaac aaagaucucg 1920 cuggugcgua cacagugcug accagcuagu guggccaggg ccugguggcc ugguggccag 1980 gaaguuucag guugaaggga aaugucgagg cuaccugcag auaugacagg ugccuugaac 2040 gcagcccauc uucaugucau caaaggucuu ccugcacuug aagcuggggc gauguuugca 2100 gucaagacca uucuuuccaa ccucuggguu cuugcaaguu gcccucaccu ugugugugga 2160 gaugcauucc aagaaugaag ccucaucuug cuacugagug ugggguucag ggaagcucuu 2220 uaggccaccu ggugaaggug cauggggagg auggagcuuc uccucagcuc cucugagcag 2280 ccaccuaugu gaucuuuaaa uccaacccca augggagaaa agggcaagaa cagucugugc 2340 ccugggacuc cuaucaggaa gcuugacagg cagcugggca ucagugcagc ugauaucguu 2400 ugaggaggga gacagaugcu uggaccuggg ugccuggcua uggagauuga ccaagcaaga 2460 ucaggagcuc cugauagcag gcgucuuuga gccuagcugg gguagaggca cugcccaucu 2520 cuucuccacc uucucuccac agaauguuug cagagcuggg caguugagga aaggacagcc 2580 ccugguuggu gccuccaaag gaagguggac uuuuuuggug gagacguuuc ugcccugggc 2640 acccuccugc ccccgauuca uaccuauggc uucuugagaa ggcucacagc uguggucuua 2700 acguagacug cagaaagaug gcaugcggcc ccuggcauuu cgccaagggu uuuauagcaa 2760 gucuccuucc uccauaggga cagcagcacc agcccugugg ggcauggagu ggaagcccag 2820 aagggcuucu gcaagcugca cagaacuggg guaagaagac aaagaguagc caccgggaga 2880 ggcuuccuuu guuacagcug ggaaagaaca guucugugaa ugcaaacacc uccugaguuu 2940 ugcaauugag aaaaugauuu ggagaacuuc ucuucuggua auuuuuauuu ugaauguuca 3000 gggccuuagu uggccccagu aauucuccuu ggaggacuug ggagaagaau uuccacaaag 3060 caaacuacua accacuagcu cuuacuggac agcgauuucu ggcuuauaag aguucucuuu 3120 gauuugcacu agcacuacga uaguguuaga uggggaaaua cugcaacaug uccaguuggc 3180 cagaucacuu uccaagggag cgauacuaag gcagacucag cuuuuuaaag augggagguc 3240 aggaggugga agugagagga gaucccaucu cacacaacac acuuccacgu aaugcagacc 3300 acacuuuucc auuuuguccu gcccucuuga gaggucauuu cucacguccu aagaaccuga 3360 ucagaaauuu uggaaggguu cuuugaaaua gcagcaguug aaacagagac acuuugccac 3420 aguguggagc agauuuucuc acugguauca cauggucuug caguuuugaa cucuucgacc 3480 gauuuguggg aguuuaugua auugcgugca augaaccuga aauuguguaa aggacaaaag 3540 accaguuuau aggguugggu uuuuuuucca acuugugaaa agcaguuuag cugcaucugu 3600 cuccccacca cccccacccc gggaggggcu uauguuacaa ggugaucaag ugaaggaaaa 3660 accugagccu aucuggcugg gaugguggaa uuaagcacaa ggucacauuc ucugugauca 3720 caugagaggg aaggugauga cuuaaauggc aggggguggg gauuaucuug gggagaggcu 3780 gaaaagcaca aaagauaguc uucccuguac guauugguga agaacgugca caaggcugga 3840 uggacuucaa cuuggaguug aguugaggca agaggauuuc uggauauuag ucacccaucu 3900 gcaagaaaaa ugcugaggcc ucgggucaag auuuugaucu gagacaugcu gaugcuucaa 3960 ggagaaauau uuucacaauc cucucuuccc ucaccagaag agaacaguac ucucuccuag 4020 aaaccucuag guaaacacau uuuauccuaa uaucgguagc auauaaugcc ccccccaaaa 4080 uaucuguuuu ccaugcaaaa aagucucaac aagaagucug uggaguugag ugguuacuuc 4140 aaagugucag gagagugaag aaauuggcca cagaagagca agaagcucuc uuaagaaaag 4200 ggaauucucu uuaaagaaac caccaccaac aacaaaacaa ccaaaaacca uguuuuaugu 4260 caaagcucug uagcacagag aauguggugu cacagauaca ucgccgagag agguuucuuu 4320 cuuucuuuuu uuuuuuuuug agacagaguc ugguucuguu ucccaggcug gagugcagug 4380 gugggaucuc agcucacugc aacauccgcc ucugggguuc aagugauucu ccugucucag 4440 ccucccaagu agcuggaauu acagggaccc gccaccacgc ccggcuaauu uuuuugugug 4500 guuuuaguag aggugggguu ucaccaucuu ggccaggcug gucuugaacu ccugaccucg 4560 ugauccaccc gccuaggccu cccaaagugu ugggauuaca ggcgugagcc acugugccca 4620 gccaaaagag aaauuucuac augaacaagg caauuucagu gucuuacagc ggccaaacca 4680 ugacgugaag aaugagauag gagacaggag aucaccauaa gcgucccuga uauagcagca 4740 cacauuuuca cguuuccacu uaaaucguuu ugcacaaagu cuugcuucgc ucagaugaga 4800 ugagauauga uuuccuagag auguaaaaau aagaaugaau guggcgcccc cuucuuccag 4860 auguaauaga aagcucugcc cuaucacaag ggggguguug aagcgccccu uguguuuuaa 4920 cuguauuuaa cugagcacaa gaugcacaag cugugguggg aaacccucag uuuaccuuug 4980 gagucuuccc ugcagaucgc agaccuguuu ccaggcugau guuucuggug uguaauugcu 5040 agcguuucug aaggguuuuc ccaauuguuu uagccuugug aaguauucuu aauuauaacu 5100 ugccuuucag cgaugguaca ugacuugauu caacguuugg uucugaacuu acacacugau 5160 gcguuuacuc aucuaacaua aucugacagg gccucagcaa gggagccaua cauuuuugua 5220 acauuuugau auguuuuaau gcaucugacu uagaucuuac ugaaauaaag cacuuuucaa 5280 agagaaaaaa aaaaaaaaaa aaaaaaa 5307 <210> 82 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> RASSF2 sense primer <400> 82 tctagatgtg tgcacacatg gtacgtgtc 29 <210> 83 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> RASSF2 antisense primer <400> 83 tctagatgta tggctccctt gctgagg 27 <210> 84 <211> 3107 <212> RNA <213> Homo sapiens VAC14 mRNA <400> 84 gacucgagua acauggccgc ugucucguga gucccgcuag ugccgggcgg gaguuguuaa 60 gcggccaggg ucaggugugc uggagcgggg uccgggcccg gguuccaggg cgaggcggcg 120 gagcguggca ggcaagccua gagcggcgug guccaugcgc cggcgccggg ggcagagcgg 180 agccgcagac uccccuggcc ccggcgcggc cccggcagcc gcgggcuaag gagucgcgag 240 guucccccag cugccaccau gaaccccgag aaggauuucg cgccgcucac gccuaacauc 300 gugcgcgccc ucaaugacaa gcuguacgaa aagcggaagg uggcagcgcu ggagaucgag 360 aagcuggucc gggaguucgu ggcccagaac aauaccgugc aaaucaagca ugugauccag 420 acccuguccc aggaguuugc ccugucucag cacccccaca gccggaaagg gggccucauc 480 ggccuggccg ccugcuccau cgcacugggc aaggacucag ggcucuaccu gaaggagcug 540 aucgagccag ugcugaccug cuucaaugau gcagacagca ggcugcgcua cuaugccugc 600 gaggcccucu acaacaucgu caagguggcc cggggcgcug ugcugcccca cuucaacgug 660 cucuuugacg ggcugagcaa gcuggcagcc gacccagacc ccaaugugaa aagcggaucu 720 gagcuccuag accgccuuuu aaaggacauu gugacugaga gcaacaaguu ugaccuggug 780 agcuucaucc ccuuguugcg agagaggauu uacuccaaca accaguaugc ccggcaguuc 840 aucaucuccu ggauccuggu ucuggagucg gugccagaca uuaaccugcu ggauuaccug 900 ccggagaucc uggauggacu cuuccagauc cugggugaca auggcaaaga gauucgcaaa 960 augugugagg uuguucuugg agaauucuua aaagaaauua agaagaaccc cuccagugug 1020 aaguuugcug agauggccaa cauccuggug auccacugcc agacaacaga ugaccucauc 1080 cagcugacag ccaugugcug gaugcgggag uucauccagc uggcgggccg cgucaugcug 1140 ccuuacuccu ccgggauccu gacugcuguc uugcccugcu uggccuacga ugaccgcaag 1200 aaaagcauca aagaaguggc caacgugugc aaccagagcc ugaugaagcu ggucaccccc 1260 gaggacgacg agcuggauga gcugagaccu gggcagaggc aggcagagcc caccccugac 1320 gaugcccugc caaagcagga gggcacagcc aguggagguc cagaugguuc cugugacucc 1380 agcuucagua gcggcaucag ugucuucacu gcagccagca cugaaagagc cccagugacc 1440 cuucaccucg acgggaucgu gcagguccua aacugccacc ucagugacac ggccauuggg 1500 augaugacca ggauugcagu ucucaagugg cucuaccacc ucuacaucaa aacuccucgg 1560 aagauguucc ggcacacgga cagccucuuu cccauccuac ugcagacguu aucggaugaa 1620 ucggaugagg ugauccugaa ggaccuggag gugcuggcag aaaucgcuuc cucccccgca 1680 ggccagacgg augacccagg cccccucgau ggcccugacc uccaggccag ccacucagag 1740 cuccaggugc ccaccccugg cagagccggc cuacugaaca ccucugguac caaaggcuua 1800 gaauguucuc cuucaacucc caccaugaau ucuuacuuuu auaaguucau gaucaaccuu 1860 cucaagagau ucagcagcga acggaagcuc cuggagguca gaggcccuuu caucaucagg 1920 cagcugugcc uccugcugaa ugcggagaac aucuuccacu caauggcaga cauccugcug 1980 cgggaggagg accucaaguu cgccucgacc augguccacg cccucaacac cauccugcug 2040 accuccacag agcucuucca gcuaaggaac cagcugaagg accugaagac ccuggagagc 2100 cagaaccugu ucugcugccu guaccgcucc uggugccaca acccagucac cacggugucc 2160 cucugcuucc ucacccagaa cuaccggcac gccuaugacc ucauccagaa guuuggggac 2220 cuggagguca ccguggacuu ccucgcagag guggacaagc uggugcagcu gauugagugc 2280 cccaucuuca cauaucugcg ccugcagcug cuggacguga agaacaaccc cuaccugauc 2340 aaggcccucu acggccugcu caugcuccug ccgcagagca gcgccuucca gcugcucucg 2400 caccggcucc agugcgugcc caacccugag cugcugcaga ccgaagacag ucuaaaggca 2460 gcccccaagu cccagaaagc ugacuccccu agcaucgacu acgcagagcu gcugcagcac 2520 uuugagaagg uccagaacaa gcaccuggaa gugcggcacc agcggagcgg gcguggggac 2580 caccuggacc ggaggguugu ccucugacag gccuggcacg gaggagggcc caccgagugg 2640 ucccaugaaa cacuaagggu cgucacgccc ucccgaggag cucaaggacc ugccugucag 2700 gaccagggcu gggccugcca acccagggca guguuggggc cggaggcugc ugugucugcc 2760 caagcuccuc ucagagucca guccccaggc cuccagcgcu gucagcugca cccuggcauu 2820 cucacagagc uggcugccca cccagugggg ggcuauagcc ucagagacca cucauccucu 2880 ggaaucaacc ucuuucuaau acccucuugg aaaaagagcu ugccccuccu ccagcacacu 2940 agagcucugg ccuugugugu auauguauac auacgugaac acaugccugu gugugugugu 3000 gugugugugu acuuguaugc acguaggcac cagcacaaag aucugaauga ugcaccccac 3060 ccccacccca auaaagaaau aacagaaaac ccucaaaaaa aaaaaaa 3107 <210> 85 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> VAC14 sense primer <400> 85 tctagaaaca ctaagggtcg tcacgcc 27 <210> 86 <211> 27 <212> DNA <213> Artificial Sequence <220> 223 VAC14 antisense primer <400> 86 tctagacttt gtgctggtgc ctacgtg 27 <210> 87 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> hsa-miR-200a <400> 87 uaacacuguc ugguaacgau gu 22 <210> 88 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> has-miR-200c <400> 88 uaauacugcc ggguaaugau gga 23 <210> 89 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> dme-miR-8 <400> 89 uaauacuguc agguaaagau guc 23 <210> 90 <211> 20 <212> DNA <213> Artificial Sequence <220> 223 Quentitative RT PCR sense primer <400> 90 attaaatcgg agcctctgga 20 <210> 91 <211> 15 <212> DNA <213> Artificial Sequence <220> 223 Quentitative RT PCR antisense primer <400> 91 ggctgtcgct ggcct 15 <210> 92 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Ced-12 Quentitative RT PCR sense primer <400> 92 gtgccggaaa actcattc 18 <210> 93 <211> 20 <212> DNA <213> Artificial Sequence <220> Ced-12 Quentitative RT PCR antisense primer <400> 93 gtacagctgg tgggtcatct 20 <210> 94 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> CG8445 Quentitative RT PCR sense primer <400> 94 catgaatcac ggcgg 15 <210> 95 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> CG8445 Quentitative RT PCR antisense primer <400> 95 tgaacttatc gtagttgtgg gt 22 <210> 96 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> CG12333 Quentitative RT PCR sense primer <400> 96 ggacttccga gaggcaat 18 <210> 97 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> CG12333 Quentitative RT PCR antisense primer <400> 97 atcgcagtcc agcagc 16 <210> 98 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Lap1 Quentitative RT PCR sense primer <400> 98 cacaacaagt gcaatatacg g 21 <210> 99 <211> 21 <212> DNA <213> Artificial Sequence <220> Lap1 Quentitative RT PCR antisense primer <400> 99 tgctattcaa agcatcttgg t 21 <210> 100 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> atro Quentitative RT PCR sense primer <400> 100 agcaggatac gcccaac 17 <210> 101 <211> 15 <212> DNA <213> Artificial Sequence <220> <223> atro Quentitative RT PCR antisense primer <400> 101 tgcggctacc ggact 15 <210> 102 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> ush RT PCR sense primer <400> 102 cgaattccga tgtatccaac g 21 <210> 103 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> ush RT PCR antisense primer <400> 103 ggagttgttg ttctcgtgca cc 22 <210> 104 <211> 34 <212> DNA <213> Artificial Sequence <220> Luc-ush 3'UTR sense primer <400> 104 caaccatcga cgacaactct cccggaaaat ctcc 34 <210> 105 <211> 34 <212> DNA <213> Artificial Sequence <220> Luc-ush 3'UTR antisense primer <400> 105 ggagattttc cgggagagtt gtcgtcgatg gttg 34 <210> 106 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Inr Quentitative RT PCR sense primer <400> 106 aacagtggcg gattcggtt 19 <210> 107 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Inr Quentitative RT PCR antisense primer <400> 107 tactcggagc attggaggca t 21 <210> 108 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> step Quentitative RT PCR sense primer <400> 108 caccaggaac ttcttgaatc 20 <210> 109 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> step Quentitative RT PCR antisense primer <400> 109 ttctttgtac caggatgtca 20 <210> 110 <211> 4507 <212> DNA <213> Homo sapiens FOG2 cDNA <400> 110 tcagcggcag cagccgccgc cgagatgtcc cggcgaaagc aaagcaaacc ccggcagatc 60 aaacggccgc ttgaagatgc cattgaagat gaggaagaag aatgtccatc agaggaaaca 120 gacatcatct ccaaaggaga ctttccattg gaggaaagct tttccacaga atttgggcct 180 gaaaatctga gctgcgaaga agtggaatac ttttgtaaca aaggtgatga tgaaggaatc 240 caggagacag cagaatcaga tggggacaca cagtcagaga aaccggggca acctggagtt 300 gagacagacg actgggatgg accaggagag ctggaggtgt ttcagaaaga tggggaacga 360 aaaattcaga gtcgacagca acttccagtg ggaacaacct gggggccgtt tcctgggaag 420 atggacttga ataataattc tttgaagaca aaggctcagg tcccaatggt gctgactgct 480 ggtcccaagt ggttgctgga tgtgacttgg caaggagtgg aagacaacaa aaacaactgc 540 attgtgtaca gcaaaggggg tcagctttgg tgtacaacta cgaaggccat ctctgagggt 600 gaagagctaa ttgcctttgt ggtggatttt gactcaaggc tacaagctgc cagtcagatg 660 actctcacag aagggatgta ccctgcacgc ctgctggact caattcagct gcttcctcag 720 caagctgcca tggcttctat tttgcccaca gctattgtca ataaggatat attcccttgc 780 aagtcctgtg gcatctggta tcggagtgag cggaatctgc aggcccattt gatgtactac 840 tgcagtggga ggcaaagaga agctgctccg gtgtcagagg aaaatgaaga cagtgcccat 900 cagatttcca gcctgtgccc cttcccacag tgcaccaaga gcttttcaaa tgctcgagct 960 ctagaaatgc acctgaattc acacagtgga gtgaaaatgg aagaattcct gccccctggt 1020 gctagtctaa aatgcaccgt ctgtagctac actgctgatt ccgtgatcaa ctttcaccaa 1080 cacctgttct cccatctcac tcaagctgcc ttccgatgta atcactgcca tttcggcttc 1140 cagactcaga gggagttatt gcagcaccag gagctccatg tccctagcgg caaacttccc 1200 agagaaagtg acatggaaca ctctccaagt gcaactgaag acagcttaca gccagccaca 1260 gacttattga ccagaagcga acttccccag agccaaaagg ccatgcagac taaagatgcg 1320 agctctgaca cagagctgga caagtgtgag aaaaagactc agctctttct cacgaaccag 1380 agaccagaga tacagcctac aacaaataaa caaagctttt cttacacaaa aataaagtct 1440 gagccctcta gcccaagact tgcctcatct ccagttcagc ctaatattgg gccttctttc 1500 cctgtgggcc ctttcctatc tcagttttct ttcccccaag atatcaccat ggtccctcaa 1560 gcttcagaga tcttagctaa gatgtctgaa ctggtgcatc ggcgactgag gcatggcagt 1620 agtagctacc ctcccgtcat ttacagccct ttgatgccca agggggctac ttgttttgag 1680 tgtaacataa cattcaataa tttggataat tatctagtgc acaaaaagca ttattgcagc 1740 agccgatggc agcagatggc taagtcccca gagttcccta gtgtgtcaga aaagatgcct 1800 gaagctttga gtcccaacac tggccaaacc tccataaacc ttctcaaccc agctgctcat 1860 tctgctgatc ctgagaatcc acttcttcaa acatcttgca tcaattcttc cactgtctta 1920 gatttaattg ggccaaatgg gaagggccat gacaaggact tttccactca aactaagaag 1980 ctctccacct ccagtaacaa tgatgacaaa attaatggaa aacctgttga tgtgaaaaat 2040 cccagtgtcc ccttagtgga tggggaaagt gacccaaata agactacctg tgaagcttgc 2100 aacattacct tcagccggca cgaaacatac atggtccaca aacagtatta ctgtgctaca 2160 cgccacgacc ctccactgaa gaggtctgct tccaacaaag tgcctgccat gcagagaacc 2220 atgcgcacac gcaagcgcag aaagatgtat gagatgtgcc tacctgagca ggaacaaagg 2280 cctccactgg ttcagcagag atttcttgac gtagccaacc tcaataatcc ttgtacctcc 2340 actcaagaac ccacagaagg gctaggagag tgctaccacc caagatgtga tatctttcca 2400 ggaattgtct ctaaacactt ggaaacttct ctgacgatca acaagtgtgt tccagtttcc 2460 aaatgtgata ctactcattc cagtgtttcc tgcctagaga tggacgtgcc catagatctc 2520 agcaaaaagt gtttatctca gtctgagcgg acgaccacgt ctcccaaaag gctgctggac 2580 tatcacgagt gcactgtgtg caagatcagt ttcaataagg tagaaaacta tctggcccac 2640 aagcagaatt tctgcccggt tactgcacat cagcgtaatg acctgggtca actggacggc 2700 aaagtgtttc cgaatccaga aagcgaacga aacagccctg atgtcagcta cgaaagaagc 2760 ataataaaat gtgagaaaaa tgggaatttg aagcagcctt cccccaatgg aaacttattt 2820 tcatcccacc tagcaacccc gcaaggcttg aaggtcttta gtgaagctgc tcagctcatt 2880 gctacaaaag aagaaaacag acatttgttt cttccacaat gcctttaccc tggagcaata 2940 aagaaagcaa aaggagccga ccagctttct ccatattatg gaatcaagcc aagtgattat 3000 atttctggtt ctcttgtcat ccataacact gacatcgagc aaagcagaaa tgcagaaaat 3060 gaatctccta aaggccaggc ttcctcaaat gggtgtgctg cgctgaagaa agattctctg 3120 ccattgttgc ccaaaaatcg aggaatggta atagtgaatg gtggactgaa acaagatgag 3180 agacctgctg ccaacccaca gcaagagaac atttcccaga atcctcagca cgaagacgac 3240 cacaaatctc cctcgtggat ctctgagaac ccattagctg ccaatgagaa tgtctcacca 3300 ggaattccct cagcagagga acagttgtct agtatagcaa aaggtgtgaa tggttccagc 3360 caggctccaa ccagtgggaa atattgccgg ctatgtgata tccagttcaa caacctttca 3420 aactttataa ctcacaagaa gttttattgc tcatcacatg cagcagaaca tgtcaaatga 3480 actaactaaa catcagtcac ctttggtatc agtgtttagt atgttgttct aaccagtcca 3540 gaaaaaaaaa taagctgttt gaattacatc tgggcaatca ggagataatt cattatggct 3600 gagttgaaga cttaaggtgt aatttcatta cagtccatta gtaaagtgta ttattggtgc 3660 cattttcaaa aaaattaatt tattttacca gcagtattca tagctgtggt tatgttattt 3720 tttatttaaa aactttatat taaagtcatt tgtaatgtta ttgtatagtt attgtgtagc 3780 acatatggtt tgcactgtat agtagctttt aaagaaaata gtcacaatac agaaaagcat 3840 tttagaaata gcttcaaaag cacttgtgta tcttgatttt ttcttatatg ctgttgcaga 3900 tatatgtata tgctaaaata taacttgcaa agatgttcta aatacacatg ctataagttc 3960 gccttaagat ttcaattctt ggataatcag gctctgtttg cactttatat tttagcagat 4020 acagtctctt agtcactagg ctttgcattt gtatgtagct gtatgtttcc gtccattttc 4080 ttaatcctga acctgtatgt taaatgaaga tggcaatttt tttcttgtat agtacttgta 4140 ttttctttcg ctgatgcagc tctgtctcaa tttttaaacc tttgctgtta aatgcaatac 4200 tttataaaga atgaacaaaa ttactggaag cagtattgta agtaatgagg tagtattaat 4260 cagttttatc ttttgaaagg cacagtctaa atcgaaaccc taaactcaat gctgcaagta 4320 tgaatttaat tcatatataa gatctattta aatataagag tagcaatact gcacctggtg 4380 atcacaaaga taatgttcta cttctgatag aaataatttc tcaacaaatg ttgttactat 4440 gcatgtatat ggatggaata aaattccaga ttgttggaga aaaaaaaaaa aaaaaaaaaa 4500 aaaaaaa 4507 <210> 111 <211> 27 <212> DNA <213> Artificial Sequence <220> 223 FOG2 cDNA sense primer <400> 111 ggatccatgt cccggcgaaa gcaaagc 27 <210> 112 <211> 32 <212> DNA <213> Artificial Sequence <220> 223 FOG2 cDNA antisense primer <400> 112 gcggccgctc atttgacatg ttctgctgca tg 32 <210> 113 <211> 37 <212> DNA <213> Artificial Sequence <220> <223> m1 sense primer <400> 113 ttaatttatt ttaccagtga cattcatagc tgtggtt 37 <210> 114 <211> 37 <212> DNA <213> Artificial Sequence <220> M1 antisense primer <400> 114 aaccacagct atgaatgtca ctggtaaaat aaattaa 37 <210> 115 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> m2 sense primer <400> 115 acaaaattac tggaagtgac attgtaagta atgagg 36 <210> 116 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> m2 antisense primer <400> 116 cctcattact tacaatgtca cttccagtaa ttttgt 36 <210> 117 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> m3 sense primer <400> 117 ttgtaagtaa tgaggtgacg ttaatcagtt ttatct 36 <210> 118 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> m3 antisense primer <400> 118 agataaaact gattaacgtc acctcattac ttacaa 36 <210> 119 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> siFOG2 <220> <223> siFOG2 <400> 119 uuauucaagu ccaucuucct t 21 <210> 120 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> anti-siGFP <400> 120 aacaucgcca ucuaauuca 19 <210> 121 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> anti-miR-141 <400> 121 ccaucuuuac cagacagugu ua 22 <210> 122 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> anti-miR-200a <400> 122 acaucguuac cagacagugu ua 22 <210> 123 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> anti-miR-200b <400> 123 ucaucauuac caggcaguau ua 22 <210> 124 <211> 23 <212> RNA <213> Artificial Sequence <220> <223> anti-miR-c <400> 124 uccaucauua cccggcagua uua 23 <210> 125 <211> 22 <212> RNA <213> Artificial Sequence <220> <223> anti-miR-429 <400> 125 acgguuuuac cagacaguau ua 22 <210> 126 <211> 1236 <212> DNA <213> FOG2 [1-412] <400> 126 tcagcggcag cagccgccgc cgagatgtcc cggcgaaagc aaagcaaacc ccggcagatc 60 aaacggccgc ttgaagatgc cattgaagat gaggaagaag aatgtccatc agaggaaaca 120 gacatcatct ccaaaggaga ctttccattg gaggaaagct tttccacaga atttgggcct 180 gaaaatctga gctgcgaaga agtggaatac ttttgtaaca aaggtgatga tgaaggaatc 240 caggagacag cagaatcaga tggggacaca cagtcagaga aaccggggca acctggagtt 300 gagacagacg actgggatgg accaggagag ctggaggtgt ttcagaaaga tggggaacga 360 aaaattcaga gtcgacagca acttccagtg ggaacaacct gggggccgtt tcctgggaag 420 atggacttga ataataattc tttgaagaca aaggctcagg tcccaatggt gctgactgct 480 ggtcccaagt ggttgctgga tgtgacttgg caaggagtgg aagacaacaa aaacaactgc 540 attgtgtaca gcaaaggggg tcagctttgg tgtacaacta cgaaggccat ctctgagggt 600 gaagagctaa ttgcctttgt ggtggatttt gactcaaggc tacaagctgc cagtcagatg 660 actctcacag aagggatgta ccctgcacgc ctgctggact caattcagct gcttcctcag 720 caagctgcca tggcttctat tttgcccaca gctattgtca ataaggatat attcccttgc 780 aagtcctgtg gcatctggta tcggagtgag cggaatctgc aggcccattt gatgtactac 840 tgcagtggga ggcaaagaga agctgctccg gtgtcagagg aaaatgaaga cagtgcccat 900 cagatttcca gcctgtgccc cttcccacag tgcaccaaga gcttttcaaa tgctcgagct 960 ctagaaatgc acctgaattc acacagtgga gtgaaaatgg aagaattcct gccccctggt 1020 gctagtctaa aatgcaccgt ctgtagctac actgctgatt ccgtgatcaa ctttcaccaa 1080 cacctgttct cccatctcac tcaagctgcc ttccgatgta atcactgcca tttcggcttc 1140 cagactcaga gggagttatt gcagcaccag gagctccatg tccctagcgg caaacttccc 1200 agagaaagtg acatggaaca ctctccaagt gcaact 1236 <210> 127 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> FOG2 1-412 sense primer <400> 127 ggatccatgt cccggcgaaa gcaaagc 27 <210> 128 <211> 28 <212> DNA <213> Artificial Sequence <220> 223 FOG2 1-412 antisense primer <400> 128 gcggccgcgt ggctggctgt aagctgtc 28 <210> 129 <211> 1131 <212> DNA <213> FOG2 [413-789] <400> 129 gaagacagct tacagccagc cacagactta ttgaccagaa gcgaacttcc ccagagccaa 60 aaggccatgc agactaaaga tgcgagctct gacacagagc tggacaagtg tgagaaaaag 120 actcagctct ttctcacgaa ccagagacca gagatacagc ctacaacaaa taaacaaagc 180 ttttcttaca caaaaataaa gtctgagccc tctagcccaa gacttgcctc atctccagtt 240 cagcctaata ttgggccttc tttccctgtg ggccctttcc tatctcagtt ttctttcccc 300 caagatatca ccatggtccc tcaagcttca gagatcttag ctaagatgtc tgaactggtg 360 catcggcgac tgaggcatgg cagtagtagc taccctcccg tcatttacag ccctttgatg 420 cccaaggggg ctacttgttt tgagtgtaac ataacattca ataatttgga taattatcta 480 gtgcacaaaa agcattattg cagcagccga tggcagcaga tggctaagtc cccagagttc 540 cctagtgtgt cagaaaagat gcctgaagct ttgagtccca acactggcca aacctccata 600 aaccttctca acccagctgc tcattctgct gatcctgaga atccacttct tcaaacatct 660 tgcatcaatt cttccactgt cttagattta attgggccaa atgggaaggg ccatgacaag 720 gacttttcca ctcaaactaa gaagctctcc acctccagta acaatgatga caaaattaat 780 ggaaaacctg ttgatgtgaa aaatcccagt gtccccttag tggatgggga aagtgaccca 840 aataagacta cctgtgaagc ttgcaacatt accttcagcc ggcacgaaac atacatggtc 900 cacaaacagt attactgtgc tacacgccac gaccctccac tgaagaggtc tgcttccaac 960 aaagtgcctg ccatgcagag aaccatgcgc acacgcaagc gcagaaagat gtatgagatg 1020 tgcctacctg agcaggaaca aaggcctcca ctggttcagc agagatttct tgacgtagcc 1080 aacctcaata atccttgtac ctccactcaa gaacccacag aagggctagg a 1131 <210> 130 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> FOG2 [413-789] sense primer <400> 130 ggatcccaga cttattgacc agaag 25 <210> 131 <211> 30 <212> DNA <213> Artificial Sequence <220> 223 FOG2 [413-789] antisense primer <400> 131 gcggccgcga tatcacatct tgggtggtag 30 <210> 132 <211> 1050 <212> DNA <213> FOG2 [802-1151] <400> 132 attgtctcta aacacttgga aacttctctg acgatcaaca agtgtgttcc agtttccaaa 60 tgtgatacta ctcattccag tgtttcctgc ctagagatgg acgtgcccat agatctcagc 120 aaaaagtgtt tatctcagtc tgagcggacg accacgtctc ccaaaaggct gctggactat 180 cacgagtgca ctgtgtgcaa gatcagtttc aataaggtag aaaactatct ggcccacaag 240 cagaatttct gcccggttac tgcacatcag cgtaatgacc tgggtcaact ggacggcaaa 300 gtgtttccga atccagaaag cgaacgaaac agccctgatg tcagctacga aagaagcata 360 ataaaatgtg agaaaaatgg gaatttgaag cagccttccc ccaatggaaa cttattttca 420 tcccacctag caaccccgca aggcttgaag gtctttagtg aagctgctca gctcattgct 480 acaaaagaag aaaacagaca tttgtttctt ccacaatgcc tttaccctgg agcaataaag 540 aaagcaaaag gagccgacca gctttctcca tattatggaa tcaagccaag tgattatatt 600 tctggttctc ttgtcatcca taacactgac atcgagcaaa gcagaaatgc agaaaatgaa 660 tctcctaaag gccaggcttc ctcaaatggg tgtgctgcgc tgaagaaaga ttctctgcca 720 ttgttgccca aaaatcgagg aatggtaata gtgaatggtg gactgaaaca agatgagaga 780 cctgctgcca acccacagca agagaacatt tcccagaatc ctcagcacga agacgaccac 840 aaatctccct cgtggatctc tgagaaccca ttagctgcca atgagaatgt ctcaccagga 900 attccctcag cagaggaaca gttgtctagt atagcaaaag gtgtgaatgg ttccagccag 960 gctccaacca gtgggaaata ttgccggcta tgtgatatcc agttcaacaa cctttcaaac 1020 tttataactc acaagaagtt ttattgctca 1050 <210> 133 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> FOG2 [802-1151] sense primer <133> 133 ggatccctct gacgatcaac aagtg 25 <210> 134 <211> 32 <212> DNA <213> Artificial Sequence <220> 223 FOG2 [802-1151] antisense primer <400> 134 gcggccgctc atttgacatg ttctgctgca tg 32 <210> 135 <211> 1518 <212> DNA <213> FOG2 [1-506] <400> 135 tcagcggcag cagccgccgc cgagatgtcc cggcgaaagc aaagcaaacc ccggcagatc 60 aaacggccgc ttgaagatgc cattgaagat gaggaagaag aatgtccatc agaggaaaca 120 gacatcatct ccaaaggaga ctttccattg gaggaaagct tttccacaga atttgggcct 180 gaaaatctga gctgcgaaga agtggaatac ttttgtaaca aaggtgatga tgaaggaatc 240 caggagacag cagaatcaga tggggacaca cagtcagaga aaccggggca acctggagtt 300 gagacagacg actgggatgg accaggagag ctggaggtgt ttcagaaaga tggggaacga 360 aaaattcaga gtcgacagca acttccagtg ggaacaacct gggggccgtt tcctgggaag 420 atggacttga ataataattc tttgaagaca aaggctcagg tcccaatggt gctgactgct 480 ggtcccaagt ggttgctgga tgtgacttgg caaggagtgg aagacaacaa aaacaactgc 540 attgtgtaca gcaaaggggg tcagctttgg tgtacaacta cgaaggccat ctctgagggt 600 gaagagctaa ttgcctttgt ggtggatttt gactcaaggc tacaagctgc cagtcagatg 660 actctcacag aagggatgta ccctgcacgc ctgctggact caattcagct gcttcctcag 720 caagctgcca tggcttctat tttgcccaca gctattgtca ataaggatat attcccttgc 780 aagtcctgtg gcatctggta tcggagtgag cggaatctgc aggcccattt gatgtactac 840 tgcagtggga ggcaaagaga agctgctccg gtgtcagagg aaaatgaaga cagtgcccat 900 cagatttcca gcctgtgccc cttcccacag tgcaccaaga gcttttcaaa tgctcgagct 960 ctagaaatgc acctgaattc acacagtgga gtgaaaatgg aagaattcct gccccctggt 1020 gctagtctaa aatgcaccgt ctgtagctac actgctgatt ccgtgatcaa ctttcaccaa 1080 cacctgttct cccatctcac tcaagctgcc ttccgatgta atcactgcca tttcggcttc 1140 cagactcaga gggagttatt gcagcaccag gagctccatg tccctagcgg caaacttccc 1200 agagaaagtg acatggaaca ctctccaagt gcaactgaag acagcttaca gccagccaca 1260 gacttattga ccagaagcga acttccccag agccaaaagg ccatgcagac taaagatgcg 1320 agctctgaca cagagctgga caagtgtgag aaaaagactc agctctttct cacgaaccag 1380 agaccagaga tacagcctac aacaaataaa caaagctttt cttacacaaa aataaagtct 1440 gagccctcta gcccaagact tgcctcatct ccagttcagc ctaatattgg gccttctttc 1500 cctgtgggcc ctttccta 1518 <210> 136 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> FOG2 [1-506] sense primer <400> 136 ggatccatgt cccggcgaaa gcaaagc 27 <210> 137 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> FOG2 [1-506] antisense primer <400> 137 gcggccgctc atttgacatg ttctgctgca tg 32 <210> 138 <211> 2367 <212> DNA <213> FOG2 [1-789] <400> 138 tcagcggcag cagccgccgc cgagatgtcc cggcgaaagc aaagcaaacc ccggcagatc 60 aaacggccgc ttgaagatgc cattgaagat gaggaagaag aatgtccatc agaggaaaca 120 gacatcatct ccaaaggaga ctttccattg gaggaaagct tttccacaga atttgggcct 180 gaaaatctga gctgcgaaga agtggaatac ttttgtaaca aaggtgatga tgaaggaatc 240 caggagacag cagaatcaga tggggacaca cagtcagaga aaccggggca acctggagtt 300 gagacagacg actgggatgg accaggagag ctggaggtgt ttcagaaaga tggggaacga 360 aaaattcaga gtcgacagca acttccagtg ggaacaacct gggggccgtt tcctgggaag 420 atggacttga ataataattc tttgaagaca aaggctcagg tcccaatggt gctgactgct 480 ggtcccaagt ggttgctgga tgtgacttgg caaggagtgg aagacaacaa aaacaactgc 540 attgtgtaca gcaaaggggg tcagctttgg tgtacaacta cgaaggccat ctctgagggt 600 gaagagctaa ttgcctttgt ggtggatttt gactcaaggc tacaagctgc cagtcagatg 660 actctcacag aagggatgta ccctgcacgc ctgctggact caattcagct gcttcctcag 720 caagctgcca tggcttctat tttgcccaca gctattgtca ataaggatat attcccttgc 780 aagtcctgtg gcatctggta tcggagtgag cggaatctgc aggcccattt gatgtactac 840 tgcagtggga ggcaaagaga agctgctccg gtgtcagagg aaaatgaaga cagtgcccat 900 cagatttcca gcctgtgccc cttcccacag tgcaccaaga gcttttcaaa tgctcgagct 960 ctagaaatgc acctgaattc acacagtgga gtgaaaatgg aagaattcct gccccctggt 1020 gctagtctaa aatgcaccgt ctgtagctac actgctgatt ccgtgatcaa ctttcaccaa 1080 cacctgttct cccatctcac tcaagctgcc ttccgatgta atcactgcca tttcggcttc 1140 cagactcaga gggagttatt gcagcaccag gagctccatg tccctagcgg caaacttccc 1200 agagaaagtg acatggaaca ctctccaagt gcaactgaag acagcttaca gccagccaca 1260 gacttattga ccagaagcga acttccccag agccaaaagg ccatgcagac taaagatgcg 1320 agctctgaca cagagctgga caagtgtgag aaaaagactc agctctttct cacgaaccag 1380 agaccagaga tacagcctac aacaaataaa caaagctttt cttacacaaa aataaagtct 1440 gagccctcta gcccaagact tgcctcatct ccagttcagc ctaatattgg gccttctttc 1500 cctgtgggcc ctttcctatc tcagttttct ttcccccaag atatcaccat ggtccctcaa 1560 gcttcagaga tcttagctaa gatgtctgaa ctggtgcatc ggcgactgag gcatggcagt 1620 agtagctacc ctcccgtcat ttacagccct ttgatgccca agggggctac ttgttttgag 1680 tgtaacataa cattcaataa tttggataat tatctagtgc acaaaaagca ttattgcagc 1740 agccgatggc agcagatggc taagtcccca gagttcccta gtgtgtcaga aaagatgcct 1800 gaagctttga gtcccaacac tggccaaacc tccataaacc ttctcaaccc agctgctcat 1860 tctgctgatc ctgagaatcc acttcttcaa acatcttgca tcaattcttc cactgtctta 1920 gatttaattg ggccaaatgg gaagggccat gacaaggact tttccactca aactaagaag 1980 ctctccacct ccagtaacaa tgatgacaaa attaatggaa aacctgttga tgtgaaaaat 2040 cccagtgtcc ccttagtgga tggggaaagt gacccaaata agactacctg tgaagcttgc 2100 aacattacct tcagccggca cgaaacatac atggtccaca aacagtatta ctgtgctaca 2160 cgccacgacc ctccactgaa gaggtctgct tccaacaaag tgcctgccat gcagagaacc 2220 atgcgcacac gcaagcgcag aaagatgtat gagatgtgcc tacctgagca ggaacaaagg 2280 cctccactgg ttcagcagag atttcttgac gtagccaacc tcaataatcc ttgtacctcc 2340 actcaagaac ccacagaagg gctagga 2367 <139> <211> 27 <212> DNA <213> Artificial Sequence <220> <223> FOG2 [1-789] sense primer <400> 139 ggatccatgt cccggcgaaa gcaaagc 27 <210> 140 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> FOG2 [1-789] antisense priemr <400> 140 gcggccgctc atttgacatg ttctgctgca tg 32  

Claims (28)

1) treating the test compound to a cell line expressing miR-200 family miRNA or miR-8 miRNA; 2) measuring the expression level or activity of FOG2 or USH protein in the cells of step 1) treated; And, 3) A method for screening an insulin signaling regulatory substance comprising the step of selecting a test compound in which the expression or activity of the FOG2 or USH protein is changed in the cells of step 1) compared to the control group. The method of claim 1, wherein the cell line expressing the miR-200 family miRNA is a human cell line derived from heart, brain, testes, liver, lung and skeletal muscle tissue. The method of claim 1, wherein the cell line expressing miR-8 miRNA is a Drosophila cell line derived from adipose tissue. The method of claim 1, wherein the test compound is any one selected from the group consisting of natural compounds, synthetic compounds, RNA, DNA, polypeptides, enzymes, proteins, ligands, antibodies, antigens, metabolites of bacteria or fungi and bioactive molecules. Method characterized in that. According to claim 1, wherein the expression level of the FOG2 or USH protein of step 2) is measured by any one method selected from the group consisting of Western blot, immunostaining, fluorescence staining and reporter assay (reporter assay). How to. According to claim 1, wherein the activity of the FOG2 or USH protein of step 2) is Iii) measuring the activity of the p85 / p110 / IRS-1 complex; Ii) measuring the expression level of FOXO mRNA or protein; And, 측정 is measured by any one method selected from the group consisting of measuring cell growth and proliferation. 7. The method of claim 6, wherein the activity of the p85 / p110 / IRS-1 complex is measured by measuring phosphorylation of AKT protein. 1) treating the test compound to a cell line expressing FOG2 or USH; 2) measuring the expression level or activity of FOG2 or USH protein in the cells of step 1); And, 3) A method for screening an insulin signaling regulatory substance comprising the step of selecting a test compound in which the expression or activity of the FOG2 or USH protein is changed in the cells of step 1) compared to the control group. 9. The method of claim 8, wherein said FOG2 expressing cell line is a human cell line derived from heart, brain, testes, liver, lung and skeletal muscle tissue. 9. The method of claim 8, wherein said USH expressing cell line is a Drosophila cell line derived from adipose tissue. The method of claim 8, wherein the test compound is any one selected from the group consisting of natural compounds, synthetic compounds, RNA, DNA, polypeptides, enzymes, proteins, ligands, antibodies, antigens, bacteria or fungi metabolites and bioactive molecules Method characterized in that. The method according to claim 8, wherein the expression level of the FOG2 or USH protein of step 2) is measured by any one method selected from the group consisting of Western blot, immunostaining, fluorescence staining and reporter assay. How to. According to claim 8, wherein the activity of the FOG2 or USH protein of step 2) is Iii) measuring the activity of the p85 / p110 / IRS-1 complex; Ii) measuring the expression level of FOXO mRNA or protein; And, 측정 is measured by any one method selected from the group consisting of measuring cell growth and proliferation. The method of claim 13, wherein the activity of the p85 / p110 / IRS-1 complex is measured by measuring phosphorylation of AKT protein. 1) contacting the FOG2 protein with the p85 protein in the presence of the test compound; 2) measuring the degree of binding between the FOG2 protein and p85 protein in the above; And, 3) A method for screening an insulin signaling regulatory substance comprising the step of selecting a test compound having a changed degree of binding between the FOG2 protein and the p85 protein in a test tube of step 2) compared to a control which is not treated with the test compound. 1) contacting the USH protein with the dp60 protein in the presence of the test compound; 2) measuring the degree of binding between the USH protein and dp60 protein in the above; And, 3) screening for an insulin signaling regulatory substance comprising the step of selecting a test compound having a changed degree of binding between the USH protein and the dp60 (Drosophila p60) protein in the test tube of step 2) compared to the control which was not treated with the test compound. Way. The method of claim 15 or 16, wherein the test compound is from the group consisting of natural compounds, synthetic compounds, RNA, DNA, polypeptides, enzymes, proteins, ligands, antibodies, antigens, metabolites of bacteria or fungi and bioactive molecules. Any one selected. The method according to claim 15 or 16, wherein the binding between the proteins of step 3) is SPR, protein chip, gel shift assay, immunoprecipitation, co-immunoassay, fluorescence immunoassay and radioimmunoassay. It is measured by any one method chosen from the group which consists of analytical methods. 1) treating the test compound to a cell line expressing the FOG2 protein; 2) measuring the binding degree of FOG2 protein and p85 protein in the cells of step 1); And, 3) A screening method of an insulin signaling regulatory substance comprising the step of selecting a test compound having a change in the binding degree of the FOG2 protein and p85 protein in the cells of step 1) compared to the control that was not treated with the test compound. 20. The method of claim 19, wherein said FOG2 expressing cell line is a human cell line derived from heart, brain, testes, liver, lung, and skeletal muscle tissue. 20. The method of claim 19, wherein the binding of the FOG2 protein and p85 protein in step 2) is SPR, protein chip, gel shift assay, immunoprecipitation, co-immunoprecipitation, fluorescence It is measured by any one method selected from the group consisting of immunoassay and radioimmunoassay. 1) treating the test compound to a cell line expressing the USH protein; 2) measuring the binding degree of USH protein and dp60 protein in the cells of step 1); And, 3) A method for screening an insulin signaling regulatory substance comprising the step of selecting a test compound having a changed degree of binding of USH protein and dp60 protein in the cells of step 1) compared to a control that is not treated with the test compound. 23. The method of claim 22, wherein said USH expressing cell line is a Drosophila cell line derived from adipose tissue. The method according to claim 22, wherein the binding of the USH protein and dp60 protein in step 2) is SPR, protein chip, gel shift assay, immunoprecipitation, co-immunoprecipitation, fluorescence It is measured by any one method selected from the group consisting of immunoassay and radioimmunoassay. 23. The method of claim 19 or 22, wherein the test compound is selected from the group consisting of natural compounds, synthetic compounds, RNA, DNA, polypeptides, enzymes, proteins, ligands, antibodies, antigens, metabolites of bacteria or fungi and bioactive molecules. Which is one of the methods. Insulin containing an antisense oligonucleotide having a sequence complementary to SEQ ID NO: 42 as a substance that inhibits the expression of FOG2 protein or an antisense oligonucleotide having a sequence complementary to SEQ ID NO: 15 as an active ingredient Composition for regulating signal transmission. 27. The composition of claim 26, wherein the antisense oligonucleotide has a nucleotide sequence of SEQ ID NO: 2, 3, 4, 9, 87, 88 or 119. delete

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