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Development of gene targeting strategies for amphibian research using genome editing

Research Project

Project/Area Number 15K06802
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Laboratory animal science
Research InstitutionHiroshima University

Principal Investigator

Suzuki Ken-ichi  広島大学, 理学研究科, 特任准教授 (90363043)

Co-Investigator(Renkei-kenkyūsha) Hayashi Toshinori  鳥取大学, 医学部, 准教授 (60580925)
Project Period (FY) 2015-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2017: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywordsネッタイツメガエル / アフリカツメガエル / イベリアトゲイモリ / ゲノム編集 / ノックイン / トランスジェニック / 両生類 / 再生 / 変態 / ノックアウト
Outline of Final Research Achievements

Following completion of the genome sequences of amphibians, gene targeting techniques have become increasingly important for the further development of amphibian research in the life sciences. Recent advances in genome editing using the CRISPR-Cas system now allow for reverse genetics in the amphibians, Xenopus laevis, Xenopus tropicalis, and Pleurodeles waltl. In order to refine these amphibians as model animals for life science research, we tried to develop a highly efficient and rapid workflow for gene knock-out, gene knock-in, conditional knock-out, and safe harbor transgenic systems.

Report

(4 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • 2015 Research-status Report
  • Research Products

    (13 results)

All 2018 2017 2016 2015 Other

All Int'l Joint Research (1 results) Journal Article (8 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 8 results,  Open Access: 4 results,  Acknowledgement Compliant: 6 results) Presentation (4 results) (of which Int'l Joint Research: 3 results,  Invited: 1 results)

  • [Int'l Joint Research] シンシナティ大学(米国)

    • Related Report
      2017 Annual Research Report
  • [Journal Article] Functional analysis of thyroid hormone receptor beta in Xenopus tropicalis founders using CRISPR-Cas2017

    • Author(s)
      Sakane Y, Iida M, Hasebe T, Fujii S, Buchholz DR, Ishizuya-Oka A, Yamamoto T, Suzuki KT
    • Journal Title

      Biology Open

      Volume: 7 Pages: 1-9

    • DOI

      10.1242/bio.030338

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Reactivation of larval keratin gene (krt62.L) in blastema epithelium during Xenopus froglet limb regeneration.2017

    • Author(s)
      Satoh A, Mitogawa K, Saito N, Suzuki M, Suzuki KT, Ochi H, Makanae A.
    • Journal Title

      Developmental Biology

      Volume: 432 Issue: 2 Pages: 265-272

    • DOI

      10.1016/j.ydbio.2017.10.015

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] A Simple Protocol for Loss-of-Function Analysis in Xenopus tropicalis founders using the CRISPR-Cas System2017

    • Author(s)
      Sakane Y, Suzuki K. T, Yamamoto T.
    • Journal Title

      Methods Mol Biol

      Volume: inpress

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Clustered Xenopus keratin genes: A genomic, transcriptomic, and proteomic analysis.2017

    • Author(s)
      Suzuki KT, Suzuki M, Shigeta M, Fortriede JD, Takahashi S, Mawaribuchi S, Yamamoto T, Taira M, Fukui A.
    • Journal Title

      Developmental Biology

      Volume: inpress

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Rapid and efficient analysis of gene function using CRISPR-Cas9 in Xenopus tropicalis founders2016

    • Author(s)
      Shigeta M, Sakane Y, Iida M, Suzuki M, Kashiwagi K, Kashiwagi A, Fujii S, Yamamoto K, Suzuki K
    • Journal Title

      Genes to Cells

      Volume: in press

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Tracking dynamics of histone H3 lysine 9 acetylation in Xenopus laevis tail regeneration2016

    • Author(s)
      Suzuki M, Takagi C, Miura S, Sakane Y, Sakuma T, Sakamoto N, Endo T, Kamei Y, Kimura H, Yamamoto T, Ueno N, Suzuki K
    • Journal Title

      Genes to Cells

      Volume: Vol.21 (4) Issue: 4 Pages: 358-369

    • DOI

      10.1111/gtc.12349

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCH systems.2016

    • Author(s)
      Sakuma T, Nakade S, Sakane Y, Suzuki K, Yamamoto T.
    • Journal Title

      Nature Protocols

      Volume: 11 Issue: 1 Pages: 118-113

    • DOI

      10.1038/nprot.2015.140

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] The Expression of TALEN before Fertilization Provides a Rapid Knock-Out Phenotype in Xenopus laevis Founder Embryos.2015

    • Author(s)
      Miyamoto K, Suzuki K, Suzuki M, Sakane Y, Sakuma T, Herberg S, Simeone A, Simpson D, Jullien J, Yamamoto T, Gurdon JB.
    • Journal Title

      PLOS ONE

      Volume: 10(11) Issue: 11 Pages: e0142946-e0142946

    • DOI

      10.1371/journal.pone.0142946

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Presentation] ネッタイツメガエルにおけるゲノム編集2018

    • Author(s)
      鈴木賢一
    • Organizer
      平成29年度NBRPネッタイツメガエル技術講習会
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] A streamlined workflow for rapid and efficient gene disruption by CRISPR-Cas9 in Xenopus tropicalis founders2017

    • Author(s)
      Shigeta M, Sakane Y, Iida M, Suzuki M, Kashiwagi K, Kashiwagi A, Fujii S, Yamamoto T and Suzuki K
    • Organizer
      Joint Meeting of the German and Japanese Societies of Developmental Biologists
    • Place of Presentation
      Kiel, Germany
    • Year and Date
      2017-03-17
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] Establishment of CRISPR-Cas9-based genome editing in Xenopus tropicalis for analysis of disease-related genes2017

    • Author(s)
      Sakane Y, Kashiwagi K, Kashiwagi A, Yamamoto T, Suzuki KT
    • Organizer
      The 8th Aquatic Animal Models of Human Disease Conference
    • Place of Presentation
      Birmingham, AL, USA
    • Year and Date
      2017-01-09
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] Clustered Xenopus keratin genes: A genomic, transcriptomic, and proteomic analysis2016

    • Author(s)
      Suzuki M, Suzuki KT, Shigeta M, Takahashi S, Mawaribuchi S, Yamamoto T, Taira M, Fukui A
    • Organizer
      第22回国際動物学会/第87回日本動物学会合同大会
    • Place of Presentation
      Okinawa, Japan
    • Year and Date
      2016-11-14
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research

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Published: 2015-04-16   Modified: 2022-06-06  

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