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Efficient chimeric mouse production technology using germ cell-deficient embryos

Research Project

Project/Area Number 17K15359
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Animal production science
Research InstitutionUniversity of Yamanashi

Principal Investigator

Nagatomo Hiroaki  山梨大学, 大学院総合研究部, 講師 (30746813)

Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2018: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywords発生率 / 遺伝資源 / 発生工学 / ゲノム編集 / マウス胚 / 生殖細胞 / 顕微操作 / 始原生殖細胞 / 胚移植 / キメラ個体 / 発生 / キメラ技術 / 胚操作
Outline of Final Research Achievements

The technology to leave a target gene for the next generation is important for medicine, livestock production, and the maintenance of endangered species. Therefore, in this study, we aimed to improve the contribution rate of target cells to germ cells compared with normal chimera by production germ cell-deficient embryos directly with fertilized eggs. It was shown that by injecting four types of Prdm14-targeted gRNA into fertilized eggs, germ cells can be efficiently deleted in about 60% of the embryos. Although chimeric mice were actually produced, the contribution to germ cells was not significantly improved as compared with the chimera using normal embryos. In addition, in this study, we developed an embryo culture technology using agarose capsules and showed that it is useful for cryopreservation.

Academic Significance and Societal Importance of the Research Achievements

目的遺伝資源を生産する方法として、キメラ技術がある。受精卵に、別の個体の受精卵やES細胞、iPS細胞などを注入、あるいは集合培養によりキメラ胚を作出することで、生まれた個体の精子や卵子は注入された受精卵やiPS細胞の遺伝情報を受け継ぐことが可能である。しかしながら、キメラ個体の場合、生殖細胞まで寄与する割合はあまり高くない場合が多い。そこで本研究で実施した手法をさらに洗練することでより効率的な遺伝資源の増産が可能となる。また、本研究で開発したアガロースカプセルを用いることで、産仔率や耐凍性が向上するため、少ない資源でも次世代へつなぐ一助となる。

Report

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

    (14 results)

All 2020 2018 2017

All Journal Article (7 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 7 results,  Open Access: 4 results,  Acknowledgement Compliant: 1 results) Presentation (7 results) (of which Int'l Joint Research: 4 results)

  • [Journal Article] Discovery of a Vertebrate-Specific Factor that Processes Flagellar Glycolytic Enolase during Motile Ciliogenesis2020

    • Author(s)
      Narita Keishi、Nagatomo Hiroaki、Kozuka-Hata Hiroko、Oyama Masaaki、Takeda Sen
    • Journal Title

      iScience

      Volume: 23 Issue: 4 Pages: 100992-100992

    • DOI

      10.1016/j.isci.2020.100992

    • URL

      https://pure.teikyo.jp/en/publications/ca2c94fe-eb98-437a-b0ce-41a7ee9865be

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Platelets play an essential role in murine lung development through Clec-2/podoplanin interaction2018

    • Author(s)
      Tsukiji Nagaharu、Inoue Osamu、Morimoto Mitsuru、Tatsumi Norifumi、Nagatomo Hiroaki、Ueta Koji、Shirai Toshiaki、Sasaki Tomoyuki、Otake Shimon、Tamura Shogo、Tachibana Toshiaki、Okabe Masataka、Hirashima Masanori、Ozaki Yukio、Suzuki-Inoue Katsue
    • Journal Title

      Blood

      Volume: 132 Issue: 11 Pages: 1167-1179

    • DOI

      10.1182/blood-2017-12-823369

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] Reciprocal regulation of TEAD4 and CCN2 for the trophectoderm development of the bovine blastocyst.2018

    • Author(s)
      Hiroki Akizawa, Ken Kobayashi , Hanako Bai, Masashi Takahashi, Shinjiro Kagawa, Hiroaki Nagatomo, Manabu Kawahara
    • Journal Title

      Reproduction

      Volume: 印刷中 Issue: 6 Pages: 563-571

    • DOI

      10.1530/rep-18-0043

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] Agarose capsules as new tools for protecting denuded mouse oocytes/embryos during handling and freezing-thawing and supporting embryonic development in vivo.2017

    • Author(s)
      Nagatomo H, Yao T, Araki Y, Mizutani E, Wakayama T.
    • Journal Title

      Scientific reports

      Volume: 7 Issue: 1 Pages: 17960-17960

    • DOI

      10.1038/s41598-017-18365-z

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Production of cloned mice using oocytes derived from ICR-outbred strain.2017

    • Author(s)
      Tanabe Y, Kuwayama H, Wakayama S, Nagatomo H, Ooga M, Kamimura S, Kishigami S, Wakayama T.
    • Journal Title

      Reproduction

      Volume: 154(6) Issue: 6 Pages: 859-866

    • DOI

      10.1530/rep-17-0372

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] Healthy offspring from freeze-dried mouse spermatozoa held on the International Space Station for 9 months2017

    • Author(s)
      Sayaka Wakayama, Yuko Kamada, Kaori Yamanaka, Takashi Kohda, Hiromi Suzuki, Toru Shimazu,
    • Journal Title

      PNAS

      Volume: 1 Issue: 23 Pages: 1-6

    • DOI

      10.1073/pnas.1701425114

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] The Number of Point Mutations In iPS Cells and ntES Cells Depends on the Method and Somatic Cell Type Employed for Their Generation.2017

    • Author(s)
      Araki R, Mizutani E, Hoki Y, Sunayama M, Wakayama S, Nagatomo H, Kasama Y, Nakamura M, Wakayama T, Abe M.
    • Journal Title

      Stem Cells

      Volume: 35 Issue: 5 Pages: 1189-1196

    • DOI

      10.1002/stem.2601

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] Development of vaginal image-based murine estrus detection system by deep le arning2020

    • Author(s)
      標智仁、兼平雅彦、神沼修、長友啓明
    • Organizer
      日本実験動物学会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 色体分配異常を起こしたマウス受精胚における異常染色体の回収と同定の試み2018

    • Author(s)
      柴﨑 郁江・鎌田 裕子・長友 啓明・大我 政敏・若山 照彦
    • Organizer
      第59回日本卵子学会
    • Related Report
      2018 Research-status Report
  • [Presentation] 種の PLC ζで活性化させたマウス卵子の発生能2018

    • Author(s)
      山本 悠ノ介・廣瀬 直樹・大 我 正敏・上 村 悟氏・長 友 啓明・若山さやか・伊 藤 潤哉・若 山 照彦
    • Organizer
      第59回日本卵子学会
    • Related Report
      2018 Research-status Report
  • [Presentation] Agarose capsules as new tools for protecting denuded mouse oocytes/embryos during handling and freezing-thawing and supporting embryonic development in vivo2017

    • Author(s)
      Hiroaki Nagatomo, Tatsuma Yao, Yasuyuki Araki, Eiji Mizutani, Teruhiko Wakayama
    • Organizer
      World Congress of Reproductive Biology 2017
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Healthy offspring from freeze-dried mouse spermatozoa held on the International Space Station for 9 months.2017

    • Author(s)
      Wakayama S, Kamada Y, Yamanaka K, Kohda T, Suzuki H, Shimazu T, Tada MN, Osada I, Nagamatsu A, Kamimura S, Nagatomo H, Mizutani E, Ishino F, Yano S, Wakayama T.
    • Organizer
      World Congress of Reproductive Biology 2017
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Developmental competence of mouse oocyte activeted by different species of PLCζ2017

    • Author(s)
      Yamamoto Y, Ooga M, Kamimura S, Nagatomo H, Wakayama S, Ito J, Wakayama T
    • Organizer
      World Congress of Reproductive Biology 2017
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Samo E, Nagatomo H, Ooga M, Kamimura S, Wakayama S, Kishigami S, Wakayama T2017

    • Author(s)
      Detailed strain specific difference on fertilization process by mouse ICSI
    • Organizer
      World Congress of Reproductive Biology 2017
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research

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

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