Production of genome-edited poultry via fertilization system

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K08083
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Integrative animal science
• Research Institution: Hokkaido University
• Principal Investigator: Mizushima Shusei 北海道大学, 理学研究院, 助教 (20515382)
• Research Collaborator: Kuroiwa Asato 北海道大学 ; Sasanami Tomohiro 静岡大学 ; Ono Tamao 信州大学
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: ウズラ受精卵 / CRISPR/Cas9 システム / 雌ゲノム

Outline of Final Research Achievements

Programmable genome editing tool was successfully adopted to establish genome-edited chicken lines. However, this method is dependent on germline-mediated chimerism mediated by primordial germ cells (PGCs), so that its practical applications have been hampered because of genetic limitation in the culture of PGCs. Thus, the aim of this study is to establish the novel method to modify avian genome. Here, I show that microinjection of CRISPR/Cas9 RNAs into 1-cell stage egg microinjected with a single sperm together with sperm-borne egg-activating factors (ICSI) is able to introduce genomic mutations on the female-specific W sex chromosome as well as autosomes and Z sex chromosomes in Japanese quail. These results indicate that combination of ICSI and CRISPR/Cas9 system could open new window for rapid generation of genome modification models in poultry.

Free Research Field

鳥類発生工学

Academic Significance and Societal Importance of the Research Achievements

研究代表者が確立してきたICSI法を基盤としたゲノム編集技術は、全ての鳥種に適用可能であるだけでなく、養鶏産業に高い経済効果が見込まれる雌家禽ゲノムへの直接的なアプローチも可能にする唯一の技術になりうる可能性がある。すなわち、鶏卵アレルゲン除去卵の大量生産をはじめとして、低アレルゲン卵でのインフルエンザワクチンの生産事業にも展開が可能になる。さらなるノックイン技術の応用は、卵白への有用タンパク質（ヒト臨床用抗体など）の大量分泌（鶏卵バイオリアクター）にも対応可能であり、医薬、飲食料品産業における新技術・新分野の構築に対しても多大な貢献が期待できる。