Establishment of the mothod to quantify methylation efficiency at a specific site

2018 Fiscal Year Final Research Report

• Project/Area Number: 17K19352
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Research Field: Molecular and Genome biology and related fields
• Research Institution: Osaka University
• Principal Investigator: Kawahara Yukio 大阪大学, 医学系研究科, 教授 (80542563)
• Project Period (FY): 2017-06-30 – 2019-03-31
• Keywords: RNA修飾 / m6A / メチル化 / エピトランスクリプトーム / ガン / 肥満

Outline of Final Research Achievements

In general, RNAs are subject to post-transcriptional modifications, which affects their fate. Among these modifications, N6-methyladenosine (m6A) is a relatively abundant modification. Given that m6A methylation is related to various diseases, such as obesity and cancer, it is important to know the dynamics of this modification and its regulatory mechanism. However, there has been no sensitive method established to measure methylation efficiency with high accuracy in a site-specific manner. In this study, we found insertion of bridged nucleic acid (BNA) into DNA probes increases the difference in melting temperature between m6A-containing RNA and unmethylated RNA. Applying this principle, we developed a dual BNA probe method, with which we successfully quantified methylation efficiency at m6A sites with high accuracy.

Free Research Field

RNA生物学

Academic Significance and Societal Importance of the Research Achievements

本研究で開発に成功したBNAプローブ法は、従来定量が不可能であった個別部位でのRNAメチル化効率の測定を可能にした。特に、リボソームRNAなどの豊富に発現しているRNA中のメチル化効率は、高感度で定量できることを示した。本研究の成果は、これまで知ることのできなかったRNAメチル化動態を計測することを可能にしただけでなく、同じ原理を利用すれば他のRNA化学修飾の定量にも応用できることを示唆している。また、その結果としてガンなどの疾患の発症病態の解明に貢献できるものと考える。