Control of fish spermatogenesis by genome editing

2018 Fiscal Year Final Research Report

• Project/Area Number: 16H04965
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Aquatic bioproduction science
• Research Institution: Hokkaido University
• Principal Investigator: FUJIMOTO TAKAFUMI 北海道大学, 水産科学研究院, 准教授 (10400003)
• Co-Investigator(Kenkyū-buntansha): 山羽 悦郎 北海道大学, 北方生物圏フィールド科学センター, 教授 (60191376)
• Research Collaborator: YAMAZAKI kyo ; NISHIHARA Hiroki
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: ゲノム編集 / 精子形成 / 不妊化

Outline of Final Research Achievements

Artificial induction of infertile fish is necessary for application of transgenic and genome edited fish for aquaculture. In this study, mutation of genes associated with sperm flagellum movement and spermiogenesis were induced by genome editing using CRISPR/Cas9 in order to develop novel male specific sterilization in zebrafish Danio rerio and dojo loach Misgurnus anguillicaudatus. We successfully induced frame shift mutations in the both genes in germ cells of F0 individuals, because the F0 males produced sperm with the mutations. The sperm showed motility as well as wild type and could fertilize to induce F1 generation. F2 generation with homozygous deficiency of spermiogenesis related gene was successfully produced by mating within F1 individuals of heterozygous mutation. But production of functional sperm was observed in the F2 generation. The unexpected functional sperm production might be caused by illegitimate translation and/or expression of orthologue genes.

Free Research Field

魚類育種遺伝学

Academic Significance and Societal Importance of the Research Achievements

遺伝子組換え技術やゲノム編集技術により作出された個体の養殖利用において，不妊化技術は生物学的封じ込めのために必須である。現在の不妊化技術は1世代限りであるため不妊形質は遺伝しないが，本研究による不妊化は雄特異的であるため，人為的な単為発生が可能な魚類では雌による変異体の継代が可能である。そのため，精子形成に関する遺伝子機能解析のモデルとして有用であるとともに，全メス生産の養殖集団に適用することで，予期せぬオスの出現による遺伝的撹乱を未然に防ぐ不妊化技術として展開できる。