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Development of highly efficient and accumulated protein expression system targeted to fish eggs

Research Project

Project/Area Number 16K14984
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Aquatic life science
Research InstitutionMie University

Principal Investigator

Tamaru Yutaka  三重大学, 生物資源学研究科, 教授 (50324554)

Research Collaborator Nakatani Hajime  名古屋大学, 工学研究科, 講師
Hori Katsutoshi  名古屋大学, 工学研究科, 教授
Project Period (FY) 2016-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2016: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Keywordsゲノム編集技術 / ゼブラフィッシュ / Crispr-Cas9 / ゲノム編集
Outline of Final Research Achievements

We attempted to construct a functional plasmid for CRISPR-Cas9 system in zebrafish. While the EF-1 alpha (zef1alpha) promoter from zebrafish developed by our laboratory was used for Cas9 protein expression, the U6 promoter newly cloned from zebrafish was used for the expression of guide RNA (gRNA). After an injection to fertilized eggs from zebrafish with the constructed plasmid, both EGFP expression downstream of Cas9 gene in the plasmid and Cas9 protein expression were observed among zebrafish 24 hours-per-fertilization (hpf) embryos. Furthermore, gRNAs designed by the vitellogenin gene in zebrafish genome were successfully expressed.

Report

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

    (5 results)

All 2017 2016

All Presentation (5 results) (of which Int'l Joint Research: 1 results)

  • [Presentation] ゼブラフィッシュ用ゲノム編集プラスミドベクターの構築2017

    • Author(s)
      滝藤 正都, 西村 渓, 田丸 浩.
    • Organizer
      第81回日本生化学会中部支部例会, 名古屋市立大学(名古屋市),2017年5月20日.
    • Related Report
      2017 Annual Research Report
  • [Presentation] ゼブラフィッシュ用ゲノム編集プラスミドベクターの構築.2017

    • Author(s)
      滝藤 正都,西村 渓,中谷 肇,堀 克敏,田丸 浩.
    • Organizer
      日本ゲノム編集学会第2回大会,千里ライフサイエンスセンター(大阪府・豊中市),2017年06月29日.
    • Related Report
      2017 Annual Research Report
  • [Presentation] ゼブラフィッシュを用いたゲノム編集用プラスミドの構築2016

    • Author(s)
      西村 渓、中谷 肇、アヴシャル-坂 恵利子、堀 克敏、田丸 浩
    • 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)
    • Related Report
      2016 Research-status Report
  • [Presentation] ゼブラフィッシュを用いたゲノム編集プラスミドの構築2016

    • Author(s)
      西村 渓、中谷 肇、アヴシャル-坂 恵利子、堀 克敏、田丸 浩
    • Organizer
      第39回日本分子生物学会年会
    • Place of Presentation
      パシフィコ横浜(神奈川県・横浜市)
    • Related Report
      2016 Research-status Report
  • [Presentation] Construction of the plasmid for genome editing using zebrafish2016

    • Author(s)
      Kei Nishimura, Hajime Nakatane, Eriko Avsar-Ban, Katsutoshi Hori, Yutaka Tamaru
    • Organizer
      iBioP 2016
    • Place of Presentation
      Haeundae Grand Hotel(韓国・プサン市)
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research

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Published: 2016-04-21   Modified: 2019-03-29  

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