Optimization of iChIP for highly sensitive identification of genome interacting molecules and application to analysis of transcriptional regulation mechanisms.
| Project/Area Number |
25830131
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Genome biology
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| Research Institution | Osaka University |
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Principal Investigator |
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| Project Period (FY) |
2013-04-01 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Keywords | ゲノム科学 / ゲノム生物学 / 遺伝子発現調節 / 発現制御 / ゲノム機能解析 / iChIP / SILAC / Pax5 / Thy28 / RNA-Seq |
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| Outline of Final Research Achievements |
Elucidation of molecular mechanisms of genome functions requires identification of molecules interacting with genomic regions of interest in vivo. In this study, we applied iChIP, one of the locus-specific chromatin immunoprecipitation technologies, to isolate a single-copy genomic locus and comprehensively identify proteins and RNAs interacting with the locus in vivo. By combining iChIP with SILAC or RNA-sequencing, we could identify proteins or RNAs interacting with the promoter region of the endogenous single-copy chicken Pax5 gene. In addition, biochemical analysis revealed that Thy28, an identified interacting protein, is functionally required for B cell-specific transcription of the Pax5 gene via recruitment of MYH9 to the Pax5 locus in DT40 chicken B cell line. Thus, iChIP is a useful tool to elucidate molecular mechanisms of genome functions.
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Report
(3 results)
Research Products
(18 results)
