Identification of the role of transcription termination and polyadenylation in ALS using multiple high-throughput sequencing analysis
| Project/Area Number |
18K06058
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 43010:Molecular biology-related
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| Research Institution | Nagoya University |
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Principal Investigator |
Masuda Akio 名古屋大学, 医学系研究科, 准教授 (10343203)
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| Project Period (FY) |
2018-04-01 – 2022-03-31
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| Project Status |
Completed (Fiscal Year 2021)
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| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Keywords | RNA結合タンパク / RNA修飾 / RNA代謝 / CLIP / tRIP / スプライシング / IDR / SRSF3 / エクソン / 転写制御 / 相分離 / 天然変性領域 / RNA-蛋白相互作用 / FUS / U1 snRNP / RNAP II / MATR3 |
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| Outline of Final Research Achievements |
We developed the tRIP method (targeted RNA immunoprecipitation), which enables identification of the protein-RNA interaction sites to a similar level of accuracy as CLIP-seq, but requires only thousands of cells. tRIP requires only 2 days to generate a cDNA library with a single RNA purification step. tRIP adopts exonuclease treatment, and abrogates tedious SDS-PAGE and membrane transfer that are essential in CLIP methodology. High sensitivity of tRIP allows us to perform serial immunoprecipitation to identify the protein-protein-RNA interaction sites starting from a subnuclear fraction.tRIP provides new insights into the regulatory mechanism of co-transcriptional RNA processing by RNA processing factors.
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| Academic Significance and Societal Importance of the Research Achievements |
従来困難であった微量検体からのRNA-蛋白結合やRNA修飾部位の解析手法の開発に成功した。従来法を大幅に上回る感度を有しながらも、手技は簡単になっており、今後、RNA-蛋白結合部位解析の標準手法となることが期待される。本手法を利用すれば、従来、少量しか入手できないために解析困難であった、初代継代培養細胞や患者検体内でのRNA代謝様式を解明できることとなり、RNA代謝の生理的意義解明や病態解明の大幅な進歩につながることが期待される。
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Report
(5 results)
Research Products
(25 results)
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[Journal Article] Ubiquitination of DNA Damage-Stalled RNAPII Promotes Transcription-Coupled Repair2020
Author(s)
Nakazawa Y, Hara Y, Oka Y, Komine O, van den Heuvel D, Guo C, Daigaku Y, Isono M, He Y, Shimada M, Kato K, Jia N, Hashimoto S, Kotani Y, Miyoshi Y, Tanaka M, Sobue A, Mitsutake N, Suganami T, Masuda A, Ohno K, Nakada S, Mashimo T, Yamanaka K, Luijsterburg MS, Ogi T
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Journal Title
Cell
Volume: 19
Issue: 6
Pages: 1228-1244
DOI
NAID
Related Report
Peer Reviewed / Int'l Joint Research
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