2023 Fiscal Year Final Research Report
Study of DNA damage and repair mechanism in porcine embryos obtained from vitrified oocytes and zygotes, and its relevance to genome editing
| Project/Area Number |
21K05912
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 42010:Animal production science-related
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| Research Institution | National Agriculture and Food Research Organization |
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Principal Investigator |
Somfai Tamas 国立研究開発法人農業・食品産業技術総合研究機構, 生物機能利用研究部門, 上級研究員 (90547720)
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| Co-Investigator(Kenkyū-buntansha) |
原口 清輝 国立研究開発法人農業・食品産業技術総合研究機構, 生物機能利用研究部門, 上級研究員 (10324576)
菊地 和弘 国立研究開発法人農業・食品産業技術総合研究機構, 生物機能利用研究部門, グループ長補佐 (20360456)
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| Project Period (FY) |
2021-04-01 – 2024-03-31
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| Keywords | ブタ卵子 / ブタ接合体 / ガラス化保存 / DNA二本鎖切断 / ゲノム編集 / OCT4 / CD46 / 胚盤胞発生 |
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| Outline of Final Research Achievements |
The purpose of this study was to correlate the degree of DNA damage and repair in vitrified porcine oocytes and fertilized eggs (zygotes) with embryonic developmental outcomes and, as feedback to the findings, to establish the technology to produce genome edited embryos from vitrified oocytes or zygotes for the first time. Vitrification significantly increased the level of DNA double-strand breaks in immature oocytes, but not in zygotes. This was highlighted with reduced embryo development in vitrified immature oocytes whereas the developmental competence of vitrified zygotes was not affected. Therefore, for the next experiments we used vitrified zygotes for the production of genome-edited porcine embryos. Genome editing was applied by knocking out the OCT4 or CD46 genes or both using CRISPRcas9 via electroporation. Over 20% of vitrified zygotes could develop to blastocyst after genome editing and the efficacy of genome editing was over 90% in all groups.
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| Free Research Field |
動物生産科学
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| Academic Significance and Societal Importance of the Research Achievements |
本研究は、卵子の超低温保存がその後の胚発生に及ぼすDNA損傷と修復のレベルに関する新たな知見を提供し、今後の家畜やヒトにおける遺伝子バンク技術の向上に貢献するものである。さらに、このプロジェクトで確立されたガラス化技術とゲノム編集技術は、ヒトの生物医学研究に使用されるゲノム編集モデルブタの生産を強化する事が可能となる。
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