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Development of mouse-non-human primate chimera analysis innovating evaluation methods of human pluripotent stem cells

Research Project

Project/Area Number 17H03568
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Laboratory animal science
Research InstitutionUniversity of Tsukuba

Principal Investigator

Sugiyama Fumihiro  筑波大学, 医学医療系, 教授 (90226481)

Co-Investigator(Kenkyū-buntansha) 村田 知弥  筑波大学, 医学医療系, 助教 (60713485)
新美 君枝  国立研究開発法人理化学研究所, 脳神経科学研究センター, 専門職研究員 (10584534)
水野 聖哉  筑波大学, 医学医療系, 助教 (10633141)
高橋 英機  国立研究開発法人理化学研究所, 脳神経科学研究センター, 副部門長 (40446521)
Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥18,460,000 (Direct Cost: ¥14,200,000、Indirect Cost: ¥4,260,000)
Fiscal Year 2019: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2018: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
Fiscal Year 2017: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000)
Keywordsレポーターノックインマウス / 原腸胚 / 三胚葉 / 多能性幹細胞 / CRISPR/Cas9 / バイシストロニック / マウス / 分化 / 内胚葉可視化マウス / 中胚葉可視化マウス / 外胚葉可視化マウス / マーモセット幹細胞 / 異種間キメラ / マーモセット / 遺伝子改変マウス / マーモセット多能性幹細胞
Outline of Final Research Achievements

Chimera formation between pluripotent stem cells (PSCs) and early embryos is the most rigorous method for assessing pluripotency of PSCs. Recently, it was revealed that human PSC injection into mouse gastrula-stage embryos resulted in efficient PSC integration and fate determination. However, it was difficult to easily analyze which germ layer tissue the PSC migrate, reside in and then differentiate. Therefore, in order to innovate the evaluation of human pluripotent stem cells, we produced a novel knock-in reporter mouse strain, MIERU (Multi-color Imaging of Embryonic Development by Reporter Units), by the CRISPR/Cas9 system and bicistronic gene expression system and then revealed that the three germ layers in MIERU mouse embryos at the gastrula stage were identified differentially.

Academic Significance and Societal Importance of the Research Achievements

マウス原腸胚において、Otx2-2A-tdTomatoマウスは外胚葉を、T-2A-TagBFPマウスは中胚葉を、Sox17-2A-EGFPマウスは内胚葉を、さらにトリプルノックインマウスは三胚葉を区別しマッピング可能であり、PSC評価のため原腸胚による異種間キメラ解析システムのツールとして有用であることを示した。またこれら各ノックインマウスは理研バイオリソース研究センターに寄託され、研究者がPSC評価や多能性研究だけでなく、in vivoにおける三胚葉分化研究等にも利用可能とした。

Report

(4 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Annual Research Report
  • 2017 Annual Research Report
  • Research Products

    (5 results)

All 2019 2018

All Journal Article (1 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (4 results) (of which Int'l Joint Research: 1 results)

  • [Journal Article] Generation of bicistronic reporter knockin mice for visualizing germ layers2019

    • Author(s)
      Suzuki H, Dinh TTH, Daitoku Y, Tanimoto Y, Kato K, Azami T, Ema M, Murata K, Mizuno S, Sugiyama F.
    • Journal Title

      Experimental Animals

      Volume: 68 Issue: 4 Pages: 499-509

    • DOI

      10.1538/expanim.19-0031

    • NAID

      130007742218

    • ISSN
      0007-5124, 1341-1357, 1881-7122
    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] 原腸胚の胚葉形成を可視化するレポーターマウスの作製2019

    • Author(s)
      鈴木 颯、Dinh TTH、大徳陽子、加藤花名子、飯島沙織、水野聖哉、杉山文博
    • Organizer
      第66回日本実験動物学会
    • Related Report
      2019 Annual Research Report
  • [Presentation] A novel creation of reporter mouse model for assessing gastrulation and three germ layer formation2018

    • Author(s)
      Hayate Suzuki, Tra Dinh Thi Houg, Seiya Mizuno and Fumihiro Sugiyama
    • Organizer
      TGSW2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 胚葉特異的ノックインレポーターマウスの作製2018

    • Author(s)
      鈴木颯, Dinh Thi Houg Tra 大徳陽子, 加藤花名子, 飯島沙織, 水野聖哉, 杉山文博
    • Organizer
      第65回日本実験動物学会総会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 原腸胚における遺伝子機能解析に有用な胚葉特異的レポーターマウスの作製2018

    • Author(s)
      鈴木颯, Tra Dinh Thi Houg, 浅見拓哉, 大徳陽子, 加藤花名子, 飯島沙織, 水野聖哉, 依馬正次, 杉山文博
    • Organizer
      日本ゲノム編集学会第3回大会
    • Related Report
      2018 Annual Research Report

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Published: 2017-04-28   Modified: 2021-02-19  

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