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Establishment of an efficient gene knock-in method using adnoviral vector and CRISPR

Research Project

Project/Area Number 15K06807
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Laboratory animal science
Research InstitutionInstitute of Physical and Chemical Research

Principal Investigator

Yoshida Tetsu  国立研究開発法人理化学研究所, 脳科学総合研究センター, 研究員 (00365438)

Co-Investigator(Kenkyū-buntansha) 三谷 幸之介  埼玉医科大学, 医学部, 教授 (10270901)
汲田 和歌子  公益財団法人実験動物中央研究所, 応用発生学研究センター, 研究員 (50549128)
徳永 暁憲  国立研究開発法人国立長寿医療研究センター, その他部局等, 室長 (70549451)
Project Period (FY) 2015-04-01 – 2018-03-31
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2017: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2015: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Keywordsマーモセット / ノックイン / ゲノム編集 / CRISPR
Outline of Final Research Achievements

Knock-in animals have been made by inducing gene targeting vector with a drug-resistant gene cassette into ES cells, followed by the drug selection. Next, chimera animals are generated using the knock-in ES cells, though the method can be applicable only for mice. Thus, we have to knock-in a gene in the genome of fertilized eggs to make knock-in marmosets. Since marmoset, a non-human primate model animal, ovulates fewer eggs than other animals, it is essential to get high rate of gene knock-in. Screening of the genes, including adenoviral coding genes, enhancing homologous recombination have been performed.

Report

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

    (7 results)

All 2018 2016

All Journal Article (1 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (6 results)

  • [Journal Article] Elavl3 is essential for the maintenance of Purkinje neuron axons2018

    • Author(s)
      Ogawa Yuki、Kakumoto Kyoko、Yoshida Tetsu、Kuwako Ken-ichiro、Miyazaki Taisuke、Yamaguchi Junji、Konno Ayumu、Hata Junichi、Uchiyama Yasuo、Hirai Hirokazu、Watanabe Masahiko、Darnell Robert B.、Okano Hideyuki、Okano Hirotaka James
    • Journal Title

      Sci Rep.

      Volume: 8 Issue: 1 Pages: 2722-2722

    • DOI

      10.1038/s41598-018-21130-5

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] Screening of the factors enhancing gene recombination using marmoset ES cells2018

    • Author(s)
      Tetsu Yoshida, Noriyuki Kishi, Hideyuki Okano
    • Organizer
      The 9th Takeda Science Foundation Symposium on PharmaScience -Genome Editing Towards Medicinal Application- Center for Learning and Innovation Takeda Pharmaceutical Company, Ltd.
    • Related Report
      2017 Annual Research Report
  • [Presentation] マーモセット受精卵を用いた遺伝子ノックイン胚の作製2018

    • Author(s)
      吉田哲、岡原純子、岡野栄之
    • Organizer
      第7回 日本マーモセット研究会大
    • Related Report
      2017 Annual Research Report
  • [Presentation] マーモセットES細胞を用いた遺伝子組換えを亢進させる因子の探索2016

    • Author(s)
      吉田哲、岸憲幸、岡野栄之
    • Organizer
      第6回 日本マーモセット研究会大会
    • Place of Presentation
      東京大学農学部弥生講堂(東京)
    • Year and Date
      2016-12-12
    • Related Report
      2016 Research-status Report
  • [Presentation] CRISPR/Cas9を用いた遺伝子ノックインマーモセットの作製2016

    • Author(s)
      吉田哲、岸憲幸、佐々木えりか、岡野栄之
    • Organizer
      第1回 日本ゲノム編集学会
    • Place of Presentation
      広島国際会議場(広島)
    • Year and Date
      2016-09-06
    • Related Report
      2016 Research-status Report
  • [Presentation] Generation of gene knock-in marmosets using CRISPR/Cas9 system2016

    • Author(s)
      Tetsu Yoshida, Noriyuki Kishi, Erika Sasaki, Hideyuki Okano
    • Organizer
      第22回 日本遺伝子細胞治療学会
    • Place of Presentation
      虎ノ門ヒルズ(東京)
    • Year and Date
      2016-07-28
    • Related Report
      2016 Research-status Report
  • [Presentation] CRISPR/Cas9を用いた遺伝子ノックインマーモセットの作製2016

    • Author(s)
      吉田哲、伊東多恵子、岸憲幸、佐々木えりか、岡野栄之
    • Organizer
      日本マーモセット研究会
    • Place of Presentation
      東京慈恵会医科大学 (東京都港区)
    • Year and Date
      2016-01-27
    • Related Report
      2015 Research-status Report

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Published: 2015-04-16   Modified: 2019-03-29  

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