Comprehensive mRNA base modification analysis reveals mechanisms of germline stem cell maintenance and differentiation

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K06628
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43060:System genome science-related
• Research Institution: Osaka University
• Principal Investigator: Kawaguchi Shinichi 大阪大学, 生命機能研究科, 助教 (40321749)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: エピトランスクリプトーム / 塩基修飾 / 生殖幹細胞 / ナノポア / Direct-mRNA-seq

Outline of Final Research Achievements

Recently, it has become clear that mRNA bases undergo modifications such as methylation, and their functions have attracted much attention. In this study, we used a fourth-generation sequencing method based on a novel nanopore device to detect sites of base modification in mRNA. As a result, we were able to identify up to about 30 candidate sites for modification per gene at the single nucleotide level. In undifferentiated germline stem cell-like cells, mRNAs encoding translation elongation factors and actin were shown to have many base modifications. Furthermore, the 3' UTR region of Men-b, one of the metabolic enzymes, has multiple base modification sites and splicing regulation, suggesting that they are related.

Free Research Field

生化学

Academic Significance and Societal Importance of the Research Achievements

遺伝子をコードする塩基配列の変化が疾患の原因となることと同様に、塩基修飾の変化に伴う疾患も知られている。これまで、mRNA上の修飾塩基を直接検出する方法はなかったが、新規なナノポア装置を用いた第４世代塩基配列決定法によって可能となってきた。この方法を用いることによって、簡便に、さまざまな細胞種における塩基修飾を測定することが出来る。今後、細胞単位での遺伝子発現情報に加えて、塩基修飾情報が蓄積され、それらを統合することによって、塩基修飾の多様な生理学的意義が明らかになると期待される。