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Establishment of a technology platform for genetic modification of fertilized eggs that enables analysis from the F0 generation

Planned Research

Project AreaA new foundation for primate developmental biology
Project/Area Number 20H05763
Research Category

Grant-in-Aid for Transformative Research Areas (B)

Allocation TypeSingle-year Grants
Review Section Transformative Research Areas, Section (III)
Research InstitutionShiga University of Medical Science

Principal Investigator

Tsukiyama Tomoyuki  滋賀医科大学, 動物生命科学研究センター, 特任准教授 (60612132)

Project Period (FY) 2020-10-02 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥38,220,000 (Direct Cost: ¥29,400,000、Indirect Cost: ¥8,820,000)
Fiscal Year 2022: ¥11,310,000 (Direct Cost: ¥8,700,000、Indirect Cost: ¥2,610,000)
Fiscal Year 2021: ¥12,350,000 (Direct Cost: ¥9,500,000、Indirect Cost: ¥2,850,000)
Fiscal Year 2020: ¥14,560,000 (Direct Cost: ¥11,200,000、Indirect Cost: ¥3,360,000)
Keywords遺伝子改変 / カニクイザル
Outline of Research at the Start

霊長類発生の理解やその制御・再構築を進めるためには、高度な発生工学技術・遺伝子改変技術の開発が必要不可欠である。
本研究では、従来法のモザイク性や低効率といった欠点を克服し、次世代型の霊長類遺伝子改変技術の基盤を構築することで領域で構築する試験管内胚発生モデルにおける機能実験を可能にすることを目的とする。
本研究班ならびに渡部班の霊長類遺伝子改変技術を、中村班、高島班で構築する試験管内霊長類胚発生モデルと組み合わせて得られる成果は、今後、他の発生生物学研究者が霊長類研究に参画する際の重要な技術基盤となることが期待され、各種細胞系譜分化の機構解明に大きく貢献できると考えられる。

Outline of Final Research Achievements

In this study, we improved the transposon vector method to achieve the goal of eliminating mosaicism in transgenic animals. We applied our originally developed piggyBac(PB) transposon-based transgenic animal generation method to cynomolgus monkeys, and succeeded in generating transgenic monkeys that express multiple fluorescent proteins throughout the body. Detailed phenotypic analysis of the monkeys revealed the expression status of the transgene in each tissue, and the position of the transgene insertion into the genome. Furthermore, using a modified PBase vector with controllable expression timing, we found the conditions under which non-mosaic animals can be produced at a high rate.

Academic Significance and Societal Importance of the Research Achievements

実験動物の遺伝子改変技術は、遺伝子機能の解明、有用動物の作出、ヒト疾患の病態解明など、生物学、医学の発展に多大な貢献をしてきた一方、小動物モデルでヒトの現象を再現するには限界もある。ヒトにおける遺伝子改変は倫理的問題について議論が尽くされておらず、ヒト胚を用いることは難しい。そこで、非ヒト霊長類胚を用いた遺伝子改変技術の確立は今後より重要度が増すと考えられる。
なお、本研究で開発したトランスポゾン転移酵素の活性制御技術は、カニクイザルへの応用のみならず、ブタやウシなどの他の大動物モデルへの応用も期待でき、生殖サイクルの長い動物種でも効率的にF0解析を行うための基盤技術となることが期待される。

Report

(4 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Annual Research Report
  • 2020 Annual Research Report
  • Research Products

    (11 results)

All 2023 2022 2021 2020

All Journal Article (8 results) (of which Peer Reviewed: 8 results,  Open Access: 8 results) Presentation (3 results) (of which Invited: 2 results)

  • [Journal Article] Induction of fetal meiotic oocytes from embryonic stem cells in cynomolgus monkeys2023

    • Author(s)
      Gyobu‐Motani Sayuri、Yabuta Yukihiro、Mizuta Ken、Katou Yoshitaka、Okamoto Ikuhiro、Kawasaki Masanori、Kitamura Ayaka、Tsukiyama Tomoyuki、Iwatani Chizuru、Tsuchiya Hideaki、Tsujimura Taro、Yamamoto Takuya、Nakamura Tomonori、Saitou Mitinori
    • Journal Title

      The EMBO Journal

      Volume: - Issue: 9

    • DOI

      10.15252/embj.2022112962

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Essential Roles of Exocyst Complex Component 3-like 2 on Cardiovascular Development in Mice2022

    • Author(s)
      Watanabe C., Shibuya H., Ichiyama Y., Okamura E., Tsukiyama-Fujii S., Tsukiyama T., Matsushita J., Azami T., Kubota Y., Ohji M., Sugiyama F., Takahashi S., Mizuno S., Tamura M., Mizutani K. & Ema M.
    • Journal Title

      Life

      Volume: 12 Issue: 11 Pages: 1730-1730

    • DOI

      10.3390/life12111730

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Conformational change of RNA-helicase DHX30 by ALS/FTD-linked FUS induces mitochondrial dysfunction and cytosolic aggregates2022

    • Author(s)
      Hikiami R, Morimura T, Ayaki T, Tsukiyama T, Morimura N, Kusui M, Wada H, Minamiyama S, Shodai A, Asada-Utsugi M, Muramatsu SI, Ueki T, Takahashi R, Urushitani M.
    • Journal Title

      Scxeintific Reports

      Volume: 12 Issue: 1 Pages: 16030-16046

    • DOI

      10.1038/s41598-022-20405-2

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] <i>Ex vivo</i> reconstitution of fetal oocyte development in humans and cynomolgus monkeys2022

    • Author(s)
      Mizuta Ken、Katou Yoshitaka、Nakakita Baku、Kishine Aoi、Nosaka Yoshiaki、Saito Saki、Iwatani Chizuru、Tsuchiya Hideaki、Kawamoto Ikuo、Nakaya Masataka、Tsukiyama Tomoyuki、Nagano Masahiro、Kojima Yoji、Nakamura Tomonori、Yabuta Yukihiro、Horie Akihito、Mandai Masaki、Ohta Hiroshi、Saitou Mitinori
    • Journal Title

      The EMBO Journal

      Volume: 41 Issue: 18

    • DOI

      10.15252/embj.2022110815

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Non-human primates as a model for human development2021

    • Author(s)
      Nakamura Tomonori、Fujiwara Kohei、Saitou Mitinori、Tsukiyama Tomoyuki
    • Journal Title

      Stem Cell Reports

      Volume: 16 Issue: 5 Pages: 1093-1103

    • DOI

      10.1016/j.stemcr.2021.03.021

    • NAID

      120007037861

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Chromosomal-scale de novo genome assemblies of Cynomolgus Macaque and Common Marmoset2021

    • Author(s)
      Jayakumar Vasanthan、Nishimura Osamu、Kadota Mitsutaka、Hirose Naoki、Sano Hiromi、Murakawa Yasuhiro、Yamamoto Yumiko、Nakaya Masataka、Tsukiyama Tomoyuki、Seita Yasunari、Nakamura Shinichiro、Kawai Jun、Sasaki Erika、Ema Masatsugu、Kuraku Shigehiro、Kawaji Hideya、Sakakibara Yasubumi
    • Journal Title

      Scientific Data

      Volume: 8 Issue: 1 Pages: 59-59

    • DOI

      10.1038/s41597-021-00935-6

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] GATA transcription factors, SOX17 and TFAP2C, drive the human germ-cell specification program2021

    • Author(s)
      Kojima Yoji、Yamashiro Chika、Murase Yusuke、Yabuta Yukihiro、Okamoto Ikuhiro、Iwatani Chizuru、Tsuchiya Hideaki、Nakaya Masataka、Tsukiyama Tomoyuki、Nakamura Tomonori、Yamamoto Takuya、Saitou Mitinori
    • Journal Title

      Life Science Alliance

      Volume: 4 Issue: 5 Pages: 202000974-202000974

    • DOI

      10.26508/lsa.202000974

    • NAID

      120006979739

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Generation of transgenic cynomolgus monkeys overexpressing the gene for amyloid-β precursor protein2020

    • Author(s)
      Seita Y, Morimura T, Watanabe N, Iwatani C, Tsuchiya H, Nakamura S, Suzuki T, Yanagisawa D, Tsukiyama T, Nakaya M, Okamura E, Muto M, Ema M, Nishimura M, Tooyama I.
    • Journal Title

      Journal of Alzheimer's Disease

      Volume: - Issue: 1 Pages: 45-60

    • DOI

      10.3233/jad-191081

    • NAID

      120006859791

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] piggyBacトランスポゾンシステムを利用したトランスジェニックカニクイザルの作出2022

    • Author(s)
      中家 雅隆, 築山 智之
    • Organizer
      日本繁殖生物学会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Generation of transgenic cynomolgus monkeys using piggyBac transposition.2021

    • Author(s)
      Tsukiyama T.
    • Organizer
      日本分子生物学会
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] Disease modeling in cynomolgus monkeys using CRISPR/Cas9.2020

    • Author(s)
      Tsukiyama T.
    • Organizer
      日本発生生物学会
    • Related Report
      2020 Annual Research Report
    • Invited

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Published: 2020-10-30   Modified: 2024-01-30  

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