Study on practical application of zinc finger nuclease for gene knockout animal production

2017 Fiscal Year Final Research Report

• Project/Area Number: 25252056
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Integrative animal science
• Research Institution: The University of Tokyo
• Principal Investigator: Naito Kunihiko 東京大学, 大学院農学生命科学研究科(農学部), 教授 (20188858)
• Co-Investigator(Renkei-kenkyūsha): Yoshizaki Goro 東京海洋大学, 海洋科学技術研究科, 教授 (70281003) ; Kikuchi Kazuhiro 独立行政法人農業生物資源研究所, 動物科学研究領域, 上級研究員 (20360456) ; Sugiura Koji 東京大学, 農学生命科学研究科, 准教授 (20595623)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Keywords: 人工ヌクレアーゼ / CRISPR / ZFN / マウス卵 / ブタ卵 / 魚類卵

Outline of Final Research Achievements

The present study was aimed to promote the practical application of artificial nucleases, which has drawn attention as a new method of gene targeting organism production, and studied various technological developments and application to livestock eggs and fish eggs.
In this study, we showed that triple KO can be prodused at a time using CRISPR/Cas system, and that large-scale mutation can be introduced. As technology development, we constructed offset nicking system which is expected to suppress nonspecific effects and increased availability of original CRISPR/Cas system of other species with different PAM sequence. For the application, we showed the availability of this system to porcine eggs, Yamame and rainbow trout eggs.

Free Research Field

生殖遺伝学