A method for analyzing dynamics of post-transcriptional modifications of small amounts of RNA

• Project/Area Number: 17H03079
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Analytical chemistry
• Research Institution: Tokyo Metropolitan University
• Principal Investigator: Taoka Masato 東京都立大学, 理学研究科, 准教授 (60271160)
• Co-Investigator(Kenkyū-buntansha): 泉 友則 山口大学, 大学院医学系研究科, 准教授 (00261694) ; 中山 洋 国立研究開発法人理化学研究所, 環境資源科学研究センター, 専任研究員 (80321793)
• Project Period (FY): 2017-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,680,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥4,080,000); Fiscal Year 2020: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2019: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2018: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2017: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
• Keywords: RNA / 転写後修飾 / 質量分析 / LC-MS

Outline of Final Research Achievements

In this study, we developed a method for purification of trace RNAs and a method for determination of their primary structures including post-transcriptional modifications (PTMs) regardless of their PTM species with high sensitivity. We also developed a high-throughput analytical method for quantifying the site-specific variation of PTM stoichiometry in femto-mol amount of RNAs. Using this method, we discovered novel PTM sites in low-molecular-weight RNAs in cells, and showed that the modification rates of several of them differ among cell lines.

Academic Significance and Societal Importance of the Research Achievements

RNAは様々な転写後修飾（PTM）を持っており、生体に重要なことが示されている。ところが、微量RNAのPTMやその動態を複数種のPTMに横断的に解析する方法は存在せず、研究が進んでいなかった。本研究はこれらの停滞を取り除く役割を持っていると考えている。