• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Analysis of the regulatory mechanism of ovulation in goats via establishing a conditional knockout animal model utilizing the genome-editing technique

Research Project

Project/Area Number 21K19187
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 42:Veterinary medical science, animal science, and related fields
Research InstitutionNagoya University

Principal Investigator

Ohkura Satoshi  名古屋大学, 生命農学研究科, 教授 (20263163)

Co-Investigator(Kenkyū-buntansha) 松山 秀一  名古屋大学, 生命農学研究科, 准教授 (50455317)
森田 康広  帯広畜産大学, 畜産学部, 准教授 (90818262)
Project Period (FY) 2021-07-09 – 2024-03-31
Project Status Completed (Fiscal Year 2023)
Budget Amount *help
¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
Fiscal Year 2023: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2021: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Keywordsゲノム編集 / キスペプチン / エレクトロポレーション / 受精卵 / GnRH / 排卵 / ヤギ / 卵胞嚢腫
Outline of Research at the Start

本研究では、ヤギをウシのモデル動物として用い、ヤギ受精卵におけるゲノム編集技術により作出したゲノム編集ヤギを用いてキスペプチン遺伝子を視床下部内局所でノックアウトし、卵胞嚢腫の病態モデルを作出することに挑戦する。ウシの受胎率は年々低下しており、畜産現場において解決すべき喫緊の課題である。受胎率低下の要因のひとつである卵胞嚢腫は、多くの卵胞が排卵することなく卵巣に存在することから、排卵を調節する視床下部神経機構の機能不全が原因と想定される。本研究では、ゲノム編集ヤギを活用した卵胞嚢腫病態モデルにより、家畜の卵胞嚢腫発症の分子機構、ひいては排卵制御中枢のメカニズムを明らかにすることをめざす。

Outline of Final Research Achievements

Our study was designed to create a goat model for follicular cysts by conditionally knocking out the kisspeptin neurons, which are pivotal in regulating ovulation, through genome editing of fertilized goat eggs using the “gene editing by electroporation of Cas9 Protein (GEEP)” method. As a first step, we attempted to establish a conditional knockout technique by creating a systemic knockout goat of the kisspeptin gene using the GEEP method. We meticulously examined and optimized electroporation conditions, leading to the successful generation of genome-edited embryos of the kisspeptin gene from internally fertilized goat eggs (1-cell stage embryos). These embryos were then transferred to recipient goats, resulting in the birth of a genome-edited male goat with a frameshift mutation with a five-nucleotide defect at the kisspeptin locus in one allele. This significant milestone represents the first-ever report of genome-edited individuals in goats.

Academic Significance and Societal Importance of the Research Achievements

本研究により、家畜におけるゲノム編集技術に欠かせない基本的な遺伝子工学および生殖工学技術を確立できたとともに、ゲノム編集技術により全身性キスペプチン遺伝子ノックアウトヤギの作出に見通しが立った。本研究の成果により、これまで報告のなかったヤギにおいてゲノム編集個体の作出技術を確立できたことは、学術的に大きな意義がある。

Report

(4 results)
  • 2023 Annual Research Report   Final Research Report ( PDF )
  • 2022 Research-status Report
  • 2021 Research-status Report
  • Research Products

    (9 results)

All 2023 2022 2021 Other

All Journal Article (1 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (6 results) (of which Int'l Joint Research: 4 results,  Invited: 2 results) Remarks (2 results)

  • [Journal Article] Estrogen increases <i>KISS1</i> expression in newly generated immortalized <i>KISS1</i>-expressing cell line derived from goat preoptic area2021

    • Author(s)
      OSHIMO Yukina、MUNETOMO Arisa、MAGATA Fumie、SUETOMI Yuta、SONODA Shuhei、TAKEUCHI Yukari、TSUKAMURA Hiroko、OHKURA Satoshi、MATSUDA Fuko
    • Journal Title

      Journal of Reproduction and Development

      Volume: 67 Issue: 1 Pages: 15-23

    • DOI

      10.1262/jrd.2020-053

    • NAID

      130007986318

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] The neuroendocrine mechanism regulating pulsatile GnRH release in the hypothalamus in goats.2023

    • Author(s)
      Ohkura, S.
    • Organizer
      Japan-Poland Joint Seminar “Cutting-edge Reproductive Physiology: Bridges to the Future of Research and Cooperation”
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] The neuroendocrine mechanism regulating pulsatile GnRH release in the hypothalamus in goats.2023

    • Author(s)
      Ohkura, S.
    • Organizer
      The 4th International Conference on Mathematics and Natural Sciences (IConMNS) 2023
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Kiss1ノックアウトヤギを用いた性腺刺激ホルモン放出ホルモンパルス発生機構の解析.2022

    • Author(s)
      近藤由梨・松山秀一・中村 翔・森田康広・稲谷結花・長尾勇佑・大蔵 聡
    • Organizer
      令和4年度東海畜産学会大会
    • Related Report
      2022 Research-status Report
  • [Presentation] Study on the method of gene delivery to the hypothalamic arcuate nucleus using goats and newly established goat kisspeptin neuron cell lines.2021

    • Author(s)
      Matsuda, F., Okuda, K., Suetomi, Y., Kobayashi, K. and Ohkura, S.
    • Organizer
      Society for Neuroscience 50th Annual Meeting (Neuroscience 2021)
    • Related Report
      2021 Research-status Report
    • Int'l Joint Research
  • [Presentation] Establishment of an immortalized KISS1-expressing cell line derived from goat preoptic area.2021

    • Author(s)
      Oshimo, Y., Munetomo, A., Magata, F., Suetomi, Y., Sonoda, S., Takeuchi, Y., Tsukamura, H., Ohkura, S. and Matsuda, F.
    • Organizer
      The 54th Annual Meeting of the Society for the Study of Reproduction
    • Related Report
      2021 Research-status Report
    • Int'l Joint Research
  • [Presentation] Kiss1ノックアウトヤギを用いたパルス状性腺刺激ホルモン分泌制御機構の解析2021

    • Author(s)
      近藤由梨・森田康広・松山秀一・稲谷結花・長尾勇佑・大蔵 聡
    • Organizer
      令和3年度愛知県農学系4機関による研究交流会
    • Related Report
      2021 Research-status Report
  • [Remarks] 名古屋大学大学院生命農学研究科動物生産科学研究室

    • URL

      https://www.agr.nagoya-u.ac.jp/~laps/

    • Related Report
      2023 Annual Research Report 2022 Research-status Report
  • [Remarks] プレスリリース:独自ゲノム編集技術を用いたゲノム編集ヤギ個体の作出に成功

    • URL

      https://www.nagoya-u.ac.jp/researchinfo/result/2022/09/post-323.html?fbclid=IwAR3Bgm3VN84MxxrQD7U8f_moMNXLLjCII-9dQ0Jb-yMTRkq0ggJAkZ61aIA

    • Related Report
      2022 Research-status Report

URL: 

Published: 2021-07-13   Modified: 2025-01-30  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi