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Developmental analysis of xenogenic germ cells using blastocyst complementation

Research Project

Project/Area Number 18H02367
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 42040:Laboratory animal science-related
Research InstitutionNational Institute for Physiological Sciences

Principal Investigator

HIRABAYASHI Masumi  生理学研究所, 行動・代謝分子解析センター, 准教授 (20353435)

Co-Investigator(Kenkyū-buntansha) 小林 俊寛  生理学研究所, 行動・代謝分子解析センター, 助教 (20587414)
Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥17,290,000 (Direct Cost: ¥13,300,000、Indirect Cost: ¥3,990,000)
Fiscal Year 2020: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2019: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
Fiscal Year 2018: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
Keywords始原生殖細胞 / 精子 / 卵子 / Prdm14遺伝子 / 生殖細胞欠損 / 異種キメラ / 胚盤胞補完法 / 多能性幹細胞 / エピブラスト / 精巣 / 卵巣 / ES/iPS細胞 / Prdm14 / 胚盤胞補完 / CRISPR/Cas9法 / ノックインラット / ノックアウトラット / H2BVenus
Outline of Final Research Achievements

Prdm14 gene-deficient rats have been developed to analyze how ES/iPS cells differentiate and develop from primordial germ cells to gametes (sperm cells or oocytes). When blastocysts from these rats were complemented with allogeneic (rat) or xenogeneic (mouse) ES/iPS cells, the presence of ES/iPS cell-derived germ cells was confirmed in the testis or ovary of the resultant chimeric offspring rats. Functional normality of these germ cells was confirmed by the ES/iPS cell-derived offspring production through natural mating (rat-rat allogeneic experimental series) or round spermatid injection (rat-mouse xenogeneic experimental series). Furthermore, the blastocyst complementation was successfully applied with genetically modified allogeneic ES/iPS cells, which could contribute to establish a rapid and efficient system for gene-modified rat production.

Academic Significance and Societal Importance of the Research Achievements

蛍光遺伝子の発現を指標にして始原生殖細胞の動向を追跡できるPrdm14遺伝子欠損ラットを用い、生殖細胞の出現時期とその後の発生過程を明らかにした。本ラットに胚盤胞補完法を適用することでES/iPS細胞由来の生殖細胞を再生できるので、様々な動物種において効率的に遺伝子改変動物を作製できるようになる。効率的な家畜動物の繁殖・生産、絶滅危惧種の保存などへの応用にも繋がりうる成果である。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Annual Research Report
  • 2018 Annual Research Report
  • Research Products

    (24 results)

All 2021 2020 2019 2018 Other

All Journal Article (8 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 8 results,  Open Access: 6 results) Presentation (9 results) (of which Invited: 2 results) Book (2 results) Remarks (5 results)

  • [Journal Article] Blastocyst complementation using Prdm14-deficient rats enables efficient germline transmission and generation of functional mouse spermatids in rats2021

    • Author(s)
      Toshihiro Kobayashi, Teppei Goto, Mami Oikawa, Makoto Sanbo, Fumika Yoshida, Reiko Terada, Naoko Niizeki, Naoyo Kajitani, Kanako Kazuki, Yasuhiro Kazuki, Shinichi Hochi, Hiromitsu Nakauchi, M. Azim Surani & Masumi Hirabayashi
    • Journal Title

      Nature Comminications

      Volume: 12 Issue: 1 Pages: 13281-10

    • DOI

      10.1038/s41467-021-21557-x

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Direct evidence that KNDy neurons maintain gonadotropin pulses and folliculogenesis as the GnRH pulse generator2021

    • Author(s)
      Nagae Mayuko、Uenoyama Yoshihisa、Okamoto Saki、Tsuchida Hitomi、Ikegami Kana、Goto Teppei、Majarune Sutisa、Nakamura Sho、Sanbo Makoto、Hirabayashi Masumi、Kobayashi Kenta、Inoue Naoko、Tsukamura Hiroko
    • Journal Title

      Proceedings of the National Academy of Sciences

      Volume: 118 Issue: 5

    • DOI

      10.1073/pnas.2009156118

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Germline development in rat revealed by visualization and deletion of Prdm142020

    • Author(s)
      Kobayashi Toshihiro、Kobayashi Hisato、Goto Teppei、Takashima Tomoya、Oikawa Mami、Ikeda Hiroki、Terada Reiko、Yoshida Fumika、Sanbo Makoto、Nakauchi Hiromitsu、Kurimoto Kazuki、Hirabayashi Masumi
    • Journal Title

      Development

      Volume: 147 Pages: 183798-183798

    • DOI

      10.1242/dev.183798

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Pluripotent stem cell-derived organogenesis in the rat model system2019

    • Author(s)
      Hirabayashi Masumi, Goto Teppei, Hochi Shinichi
    • Journal Title

      Transgenic Research

      Volume: 28 Issue: 3-4 Pages: 287-297

    • DOI

      10.1007/s11248-019-00161-2

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Humanized UGT2 and CYP3A transchromosomic rats for improved prediction of human drug metabolism2019

    • Author(s)
      Kazuki Yasuhiro、Kobayashi Kaoru、他23名
    • Journal Title

      Proceedings of the National Academy of Sciences

      Volume: 116 Issue: 8 Pages: 3072-3081

    • DOI

      10.1073/pnas.1808255116

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Generation of pluripotent stem cell-derived mouse kidneys in Sall1-targeted anephric rats2019

    • Author(s)
      Goto Teppei、Hara Hiromasa、Sanbo Makoto、Masaki Hideki、Sato Hideyuki、Yamaguchi Tomoyuki、Hochi Shinichi、Kobayashi Toshihiro、Nakauchi Hiromitsu、Hirabayashi Masumi
    • Journal Title

      Nature Communications

      Volume: 10 Issue: 1 Pages: 451-451

    • DOI

      10.1038/s41467-019-08394-9

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Contribution of rat embryonic stem cells to xenogeneic chimeras in blastocyst or 8-cell embryo injection and aggregation.2019

    • Author(s)
      Okumura H, Nakanishi A, Toyama S, Yamanoue M, Yamada K, Ukai A, Hashita T, Iwao T, Miyamoto T, Tagawa YI, Hirabayashi M, Miyoshi I, Matsunaga T.
    • Journal Title

      Xenotransplantation

      Volume: 26(1) Issue: 1

    • DOI

      10.1111/xen.12468

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] An interspecies barrier to tetraploid complementation and chimera formation2018

    • Author(s)
      Yamaguchi Tomoyuki、Sato Hideyuki、Kobayashi Toshihiro、Kato-itoh Megumi、Goto Teppei、Hara Hiromasa、Mizuno Naoaki、Yanagida Ayaka、Umino Ayumi、Hamanaka Sanae、Suchy Fabian、Masaki Hideki、Ota Yasunori、Hirabayashi Masumi、Nakauchi Hiromitsu
    • Journal Title

      Scientific Reports

      Volume: 8 Issue: 1 Pages: 15289-15298

    • DOI

      10.1038/s41598-018-33690-7

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] アデノ随伴ウイルスベクター法を介した効率的なTfap2c-T2A-tdTomatoノックインラット作製とその発現解析2020

    • Author(s)
      長江麻佑子, 及川真実,水野直彬, 岩月研祐, 三宝 誠, 中内啓光,平林真澄,小林俊寛
    • Organizer
      第67回日本実験動物学会
    • Related Report
      2020 Annual Research Report
  • [Presentation] シルクフィブロインディスクをデバイスとしてSSV法によってガラス化保存したラット膵島の腎被膜下移植2020

    • Author(s)
      岩月研祐, 山中貴寛, 根岸 淳, 手島淳輝, 玉田 靖, 平林真澄, 保地眞一
    • Organizer
      第67回日本実験動物学会
    • Related Report
      2020 Annual Research Report
  • [Presentation] Prdm14遺伝子をノックアウトしたラット性腺の免疫組織学的解析2019

    • Author(s)
      平林 真澄,後藤 哲平,三宝 誠, 保地 眞一, 中内 啓光, 小林 俊寛
    • Organizer
      第66回日本実験動物学会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Prdm14 遺伝子座に H2BVeunus をノックインしたラットにおける始原生殖細胞発生の解析2019

    • Author(s)
      小林 俊寛,小林 久人,後藤 哲平,及川 真実,三宝 誠,中内 啓光,栗本 一基,平林 真澄
    • Organizer
      第66回日本実験動物学会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Prdm14遺伝子ノックアウトによる生殖細胞欠損ラットの作製とその応用2019

    • Author(s)
      平林 真澄,後藤 哲平,三宝 誠, 保地 眞一, 中内 啓光, 小林 俊寛
    • Organizer
      第112回日本繁殖生物学会
    • Related Report
      2019 Annual Research Report
  • [Presentation] ラット発生工学技術の軌跡2019

    • Author(s)
      平林 真澄
    • Organizer
      日本実験動物協会関東支部第20回REG部会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] 異種間キメラ動物における自己寛容破綻のメカニズムの解明2019

    • Author(s)
      濱仲 早苗,後藤 哲平,平林 真澄,山口 智之,中内 啓光.
    • Organizer
      第21回日本異種移植研究会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 前核注入時のCas9 nucleaseデリバリー様式がラット胚盤胞の変異モザイク発生頻度に及ぼす影響2018

    • Author(s)
      後藤 哲平, 余郷 享子, 保地 眞一, 平林 真澄.
    • Organizer
      第65回日本実験動物学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] ヒト初期発生、とくに生殖細胞への運命決定機構の解明に向けた複合的アプローチ2018

    • Author(s)
      小林 俊寛
    • Organizer
      第21回日本異種移植研究会
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Book] Embryonic stem cells and gene manipulation in rat. (Eds. Hayman G., Smith J., Dwinell M., Shimoyama M.) Rat Genomics. Methods in Molecular Biology.2019

    • Author(s)
      Hirabayashi Masumi, Takizawa Akiko, Hochi Shinichi
    • Total Pages
      16
    • Publisher
      Humana Press, New York.
    • ISBN
      9781493995806
    • Related Report
      2019 Annual Research Report
  • [Book] Organ generation from knocked-out rat blastocysts complemented with pluripotent stem cells (Eds. Liu C and Du Y) “Microinjection: Methods and Protocols”2019

    • Author(s)
      Hirabayashi M, Hochi S
    • Total Pages
      14
    • Publisher
      Humana Press, Totowa.
    • Related Report
      2018 Annual Research Report
  • [Remarks] 生理学研究所ホームページ

    • URL

      http://www.nips.ac.jp/

    • Related Report
      2020 Annual Research Report
  • [Remarks] 行動・代謝分子解析センター 遺伝子改変動物作製室(平林研究室)ホームページ

    • URL

      http://www.nips.ac.jp/mamtg/

    • Related Report
      2020 Annual Research Report 2019 Annual Research Report
  • [Remarks] 生理学研究所ホームページ

    • URL

      https://www.nips.ac.jp/

    • Related Report
      2019 Annual Research Report
  • [Remarks] 生理学研究所

    • URL

      https://www.nips.ac.jp/

    • Related Report
      2018 Annual Research Report
  • [Remarks] 行動・代謝分子解析センター 遺伝子改変動物作製室(平林研究室)

    • URL

      http://www.nips.ac.jp/mamtg/

    • Related Report
      2018 Annual Research Report

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Published: 2018-04-23   Modified: 2022-01-27  

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