Novel method to identify targets for RNA binding protein by using RNA modification enzyme.

• Project/Area Number: 24651215
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Genome biology
• Research Institution: Osaka University
• Principal Investigator: KUMAGAI Yutaro 大阪大学, 免疫学フロンティア研究センター, 特任助教 (00528408)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2013: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: RNA結合タンパク質

Outline of Final Research Achievements

In this project, enzymatic active region of an enzyme which position-specifically 5-methylates cytosine in RNA will be fused to RNA binding protein of interest to render the enzyme target specificity. The enzyme will specifically methylate target RNA, which can be distinguished by using bisulfite sequencing. This project aims at identifying targets of RNA binding protein of interest through identification of methylated RNAs by using the method above. By optimizing reaction and expression system of the fusion enzymes, we succeeded to apply the proposed method in certain cell lines when enzyme-fusion proteins are over-expressed. We further continue to search for enzymes which can be used in primary cells, and also plan to identify targets of RNA binding proteins in cell lines.