• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Development of a new method for satiotemporal gene function by the CRISPR/Cas9 system

Research Project

Project/Area Number 26870845
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Genetics/Chromosome dynamics
Developmental biology
Research InstitutionUniversity of Yamanashi

Principal Investigator

OTA Satoshi  山梨大学, 総合研究部, 医学研究員 (60553310)

Project Period (FY) 2014-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
KeywordsCRISPR/Cas9 / ゼブラフィッシュ / ノックイン / トランスジェニック / 遺伝子破壊 / ノックアウト / Gal4-UAS / GFP / pax2a / ゲノム編集 / コンディショナルノックアウト
Outline of Final Research Achievements

In this study, we generated transgenic zebrafish that expresses nuclease Cas9 under control of the UAS promoter. However, guide RNA injection in transgenic embryos that express GFP did not induce insertion or deletion mutations at the target locus.
We also developed a new method for analyzing gene function and expression using CRISPR/Cas9. We inserted the donor vector containing a heat-shock promoter and GFP into the 5'-untranslated region of the pax2a locus. These embryos expressed GFP at the region where endogenous pax2a expresses. We found that homozygous transgenic embryos presented the pax2a mutant phenotype. Thus, targeted insertion of the reporter gene at the endogenous gene is useful for analyzing gene expression and function.

Report

(3 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • Research Products

    (18 results)

All 2015 2014 Other

All Journal Article (4 results) (of which Peer Reviewed: 4 results,  Acknowledgement Compliant: 3 results,  Open Access: 2 results) Presentation (10 results) Book (2 results) Remarks (2 results)

  • [Journal Article] Comprehensive analysis of sphingosine-1-phosphate receptor mutants during zebrafish embryogenesis.2015

    • Author(s)
      Hisano Y, Inoue A, Taimatsu K, Ota S, Ohga R, Kotani H, Muraki M, Aoki J, Kawahara A
    • Journal Title

      Genes Cells.

      Volume: 20 Issue: 8 Pages: 647-658

    • DOI

      10.1111/gtc.12259

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Precise in-frame integration of exogenous DNA mediated by CRISPR/Cas9 system in zebrafish.2015

    • Author(s)
      Hisano Y, Sakuma T, Nakade S, Ohga R, Ota S, Okamoto H, Yamamoto T, Kawahara A.
    • Journal Title

      Sci Rep.

      Volume: 5 Issue: 1 Pages: 8841-8841

    • DOI

      10.1038/srep08841

    • Related Report
      2015 Annual Research Report 2014 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Efficient Multiple Genome Modifications Induced by the crRNAs, tracrRNA and Cas9 Protein Complex in Zebrafish.2015

    • Author(s)
      Hirohito Kotani , Kiyohito Taimatsu , Rie Ohga, Satoshi Ota, Atsuo Kawahara
    • Journal Title

      PLoS One

      Volume: 10 Issue: 5 Pages: e0128319-e0128319

    • DOI

      10.1371/journal.pone.0128319

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Multiple genome modifications by the CRISPR/Cas9 system in zebrafish2014

    • Author(s)
      Satoshi Ota, Yu Hisano, Yohshiya Ikawa, Atsuo Kawahara
    • Journal Title

      Genes to Cells

      Volume: 19 Issue: 7 Pages: 555-564

    • DOI

      10.1111/gtc.12154

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Presentation] Targeted insetion of a reporter constract using CRISPR/Cas92015

    • Author(s)
      Satoshi Ota, Kiyohito Taimatsu, Rie Ohga, Atsuo Kawahara
    • Organizer
      第21回小型魚類研究会
    • Place of Presentation
      大阪大学(大阪府吹田市)
    • Year and Date
      2015-09-19
    • Related Report
      2015 Annual Research Report
  • [Presentation] Multiple genome modifications induced by a ready-to-use CRISPR/Cas9 system in zebrafish2015

    • Author(s)
      Rie Ohga, Kiyohito Taimatsu, Hirohito Kotani, Satoshi Ota, Atsuo Kawahara
    • Organizer
      第21回小型魚類研究会
    • Place of Presentation
      大阪大学(大阪府吹田市)
    • Year and Date
      2015-09-19
    • Related Report
      2015 Annual Research Report
  • [Presentation] Spatially restricted Rel/NF-kappaB activation directs dorsal-ventral patterning of the zebrafish embryo2015

    • Author(s)
      Satoshi Anai, Satoshi Ota, Shizuka Ishitani, Tohru Ishitani
    • Organizer
      第21回小型魚類研究会
    • Place of Presentation
      大阪大学(大阪府吹田市)
    • Year and Date
      2015-09-19
    • Related Report
      2015 Annual Research Report
  • [Presentation] Multiple geneme modifications induced by crRNAs, tracrRNA and Cas9 protein in zebrafish2015

    • Author(s)
      Kiyohito Taimatsu, Hirohito Kotani, Rie Ohga, Satoshi Ota, Atsuo Kawahara
    • Organizer
      第48回日本発生生物学会大会
    • Place of Presentation
      つくば国際会議場(茨城県つくば市)
    • Year and Date
      2015-06-02
    • Related Report
      2015 Annual Research Report
  • [Presentation] The NFkappaB transcription factor Rel fine-tunes dorsal-ventral pattern of zebrafish early embryos2015

    • Author(s)
      Satoshi Anai, Satoshi Ota, Shizuka Ishitani, Tohru Ishitani
    • Organizer
      第48回日本発生生物学会大会
    • Place of Presentation
      つくば国際会議場(茨城県つくば市)
    • Year and Date
      2015-06-02
    • Related Report
      2015 Annual Research Report
  • [Presentation] NF-kappa Bファミリーの転写因子c-RelはAdmpの発現制御を介して脊椎動物初期胚の背腹軸形成を制御する2014

    • Author(s)
      穴井 諭、太田 聡、石谷 太
    • Organizer
      第37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)
    • Year and Date
      2014-11-25 – 2014-11-27
    • Related Report
      2014 Research-status Report
  • [Presentation] Precise genome-editing using CRISPR/Cas9 system2014

    • Author(s)
      Yu Hisano, Satoshi Ota, Hitoshi Okamoto, Atsuo Kawahara
    • Organizer
      第20回小型魚類研究会
    • Place of Presentation
      慶應義塾大学薬学部(東京都)
    • Year and Date
      2014-09-20 – 2014-09-21
    • Related Report
      2014 Research-status Report
  • [Presentation] The NF-kappa B transcription factor c-Rel regulates zebrafish dorsal-ventral patterning2014

    • Author(s)
      Satoshi Anai, Satoshi Ota, Tohru Ishitani
    • Organizer
      第20回小型魚類研究会
    • Place of Presentation
      慶應義塾大学薬学部(東京都)
    • Year and Date
      2014-09-20 – 2014-09-21
    • Related Report
      2014 Research-status Report
  • [Presentation] CRISPR/Cas9-mediated genome modifications in zebrafish2014

    • Author(s)
      Satoshi Ota, Yu Hisano, Atsuo Kawahara
    • Organizer
      11th Internatinal Conference on Zebrafish Development and Genetics
    • Place of Presentation
      マディソン(アメリカ合衆国)
    • Year and Date
      2014-06-24 – 2014-06-28
    • Related Report
      2014 Research-status Report
  • [Presentation] CRISPR/Cas9-mediated genome modifications in zebrafish2014

    • Author(s)
      Satoshi Ota, Yu Hisano, Atsuo Kawahara
    • Organizer
      第47回日本発生生物学会
    • Place of Presentation
      ウインクあいち(愛知県名古屋市)
    • Year and Date
      2014-05-27 – 2014-05-30
    • Related Report
      2014 Research-status Report
  • [Book] 進化するゲノム編集技術 第2編 第2章 第3節「ゼブラフィッシュにおけるゲノム編集」2015

    • Author(s)
      太田聡、川原敦雄
    • Total Pages
      348
    • Publisher
      エヌティーエス
    • Related Report
      2015 Annual Research Report
  • [Book] 実験医学7月号2014

    • Author(s)
      太田 聡、川原敦雄
    • Total Pages
      137
    • Publisher
      羊土社
    • Related Report
      2014 Research-status Report
  • [Remarks] ゼブラフィッシュにおけるCRISPR/Cas9システムを用いた外来遺伝子の精巧なゲノム挿入法

    • URL

      http://www.natureasia.com/ja-jp/srep/abstracts/62928

    • Related Report
      2015 Annual Research Report
  • [Remarks] 論文紹介

    • URL

      http://www.natureasia.com/ja-jp/srep/abstracts/62928

    • Related Report
      2014 Research-status Report

URL: 

Published: 2014-04-04   Modified: 2017-05-10  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi