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

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K19187
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 42:Veterinary medical science, animal science, and related fields
• Research Institution: Nagoya 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
• 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 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.

Free Research Field

家畜繁殖学

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。