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Specific detection of miRNA by mismatch-binding protein-mediated reverse transcription

Research Project

Project/Area Number 16K18680
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Applied microbiology
Research InstitutionOsaka Medical College

Principal Investigator

Fukui Kenji  大阪医科大学, 医学部, 助教 (00466038)

Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Keywordsミスマッチ / タンパク質 / 逆転写 / miRNA / 逆転写反応 / 高度好熱菌 / PCR / X線結晶構造 / 蛋白質 / 応用微生物 / バイオテクノロジー / 核酸 / 遺伝子
Outline of Final Research Achievements

Homologous miRNAs have only a few base differences to each other. Some of them control the expression of oncogenes; therefore, analyses of their expression profiles are important for cancer diagnosis and therapeutic strategies. In this study, we aimed to establish a simple method to identify expression patterns of homologous miRNAs by highly accurate reverse transcription using mismatch recognition proteins. From the group of functionally unknown proteins derived from extremely-thermophilic bacterium, we identified proteins capable of recognizing unpaired bases consisting of DNA / RNA hybrids. It was revealed that those proteins specifically suppress reverse transcription from mispaired primers and showed that they could be used for high-precision reverse transcription.

Academic Significance and Societal Importance of the Research Achievements

癌の基礎研究、診断や治療戦略策定手順の改善は国民の福祉に関する大きな課題である。各種癌組織で特異的な発現パターンを示す相同miRNAの正確な解析は上記の課題にとって今後益々必要になると予想される。本研究では、ミスマッチ認識タンパク質を利用することで、相同miRNA発現パターンの簡便な解析方法の構築を目指した。まず、複数のミスマッチ認識タンパク質を新たに同定する事に成功した。さらに、これらのタンパク質が逆転写反応の高精度化に有効であることを示した。今後、この手法をさらに発展させることが出来れば癌に関する基礎研究や診断手法の前進に貢献できる可能性がある。

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • 2016 Research-status Report
  • Research Products

    (3 results)

All 2019 2018 Other

All Journal Article (1 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (1 results) Remarks (1 results)

  • [Journal Article] Biochemical characterization of mismatch-binding protein MutS1 and nicking endonuclease MutL from a euryarchaeon Methanosaeta thermophila.2019

    • Author(s)
      Ai Minobe, Kenji Fukui, Hitomi Yonezu, Koki Ohshita, Saki Mizobuchi, Takashi Morisawa, Yuichi Hakumai, Takato Yano, Makoto Ashiuchi, Taisuke Wakamatsu
    • Journal Title

      DNA Repair

      Volume: 75 Pages: 29-38

    • DOI

      10.1016/j.dnarep.2019.01.005

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] Methanosaeta thermophila 由来ミスマッチ結合タンパク質 MutS1 及びニッキングヌクレアーゼ MutL 生化学的機能解析2018

    • Author(s)
      美濃部 亜衣、福井 健二、米津 ひとみ、大下 紘貴、溝渕 早紀、森澤 高至、白米 優一、矢野 貴人、芦内 誠、若松 泰介
    • Organizer
      第19回 極限環境生物学会 年会
    • Related Report
      2018 Annual Research Report
  • [Remarks] 核酸が形成するミスマッチに関わるタンパク質

    • URL

      http://www.osaka-med.ac.jp/deps/med/staff/fukui/

    • Related Report
      2016 Research-status Report

URL: 

Published: 2016-04-21   Modified: 2020-03-30  

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