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Understanding the origin and nature of meiosis - approaching from analysis of parthenogenetic oogenesis in Daphnia pulex

Research Project

Project/Area Number 16K18587
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Genetics/Chromosome dynamics
Research InstitutionHokkaido University (2017-2018)
Iwate Medical University (2016)

Principal Investigator

Hiruta Chizue  北海道大学, 理学研究院, 研究院研究員 (20723018)

Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywordsミジンコ / 単為生殖 / 減数分裂 / ゲノム編集 / 生殖細胞形成 / ノックアウト / ノックイン
Outline of Final Research Achievements

In order to reveal the molecular mechanism of the parthenogenetic oogenesis in the water flea Daphnia pulex, the establishment of genome editing technique is needed. In this study, I conducted microinjection of CRISPR/Cas9 ribonucleoprotein complexes (Cas9 RNPs) into early embryos and examined the possibility of using CRISPR/Cas9 to edit the genome. As a result, I have succeeded the development of targeted gene disruption (knock-out) by using Cas9 RNPs in D. pulex. Moreover, I advance the research in the development of knock-in system by using both TALEN and CRISPR/Cas9.

Academic Significance and Societal Importance of the Research Achievements

単為生殖は、卵が受精することなく単独で発生して個体が生じる現象である。単為生殖は卵を形成する分裂である「減数分裂」を変化させて実現されているため、単為生殖の機構の解明は減数分裂の機構の理解につながる。本研究では、ミジンコを用いて単為生殖の機構を明らかにすることを目指し、ゲノム編集法のひとつであるCRISPR/Cas9による標的遺伝子の破壊法を確立した。これにより、単為生殖に関わる遺伝子の機能を解析することが可能になったので、今後の研究進展が期待される。

Report

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

    (5 results)

All 2018 2016 Other

All Journal Article (2 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 2 results,  Open Access: 1 results) Presentation (2 results) Remarks (1 results)

  • [Journal Article] Targeted gene disruption by use of CRISPR/Cas9 ribonucleoprotein complexes in the water flea Daphnia pulex2018

    • Author(s)
      Hiruta Chizue、Kakui Keiichi、Tollefsen Knut E.、Iguchi Taisen
    • Journal Title

      Genes to Cells

      Volume: 23 Issue: 6 Pages: 494-502

    • DOI

      10.1111/gtc.12589

    • NAID

      120006636176

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Targeted gene disruption by use of CRISPR/Cas9 ribonucleoprotein complexes in the water flea <i>Daphnia pulex</i>2018

    • Author(s)
      Chizue Hiruta, Keiichi Kakui, Knut E. Tollefsen, Taisen Iguchi
    • Journal Title

      Genes to Cells

      Volume: 印刷中

    • NAID

      120006636176

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Int'l Joint Research
  • [Presentation] ミジンコのゲノム編集基盤の確立へ向けた現状と課題2018

    • Author(s)
      蛭田千鶴江, 角井敬知, 井口泰泉
    • Organizer
      日本動物学会第89回大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] TAL-PITCh法を用いたノックイン法の確立へ向けて[Approach to establishing a knock-in system by using TAL-PITCh in the water flea Daphnia pulex]2016

    • Author(s)
      蛭田千鶴江, 佐久間哲史, 荻野由紀子, 山本卓, 井口泰泉
    • Organizer
      日本ゲノム編集学会第1回大会
    • Place of Presentation
      広島国際会議場(広島県・広島市)
    • Year and Date
      2016-09-06
    • Related Report
      2016 Research-status Report
  • [Remarks] 蛭田千鶴江 Chizue Hiruta

    • URL

      https://sites.google.com/site/chizuehiruta/

    • Related Report
      2018 Annual Research Report 2017 Research-status Report

URL: 

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

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