Application of CRISPR/Cas9 system for gene manipulation in mice

2016 Fiscal Year Final Research Report

• Project/Area Number: 25250014
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Laboratory animal science
• Research Institution: Osaka University
• Principal Investigator: IKAWA MASAHITO 大阪大学, 微生物病研究所, 教授 (20304066)
• Co-Investigator(Renkei-kenkyūsha): ISOTANI Ayako 奈良先端科学技術大学院大学, 准教授 (20444523) ; FUJIHARA Yoshitaka 大阪大学, 微生物病研究所, 助教 (70578848) ; MIYATA Haruhiko 大阪大学, 微生物病研究所, 助教 (50604732) ; SATOUH Yuhkoh 大阪大学, 微生物病研究所, 助教 (40545571)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Keywords: ゲノム編集 / 生殖不全 / 遺伝子破壊 / 点変異 / ES細胞

Outline of Final Research Achievements

We have introduced CRISPR/Cas9 system to generate genome edited mice. First, we prepared pCAG-EGxxFP plasmid to assess gRNA cleavage efficiency. Next, we injected gRNA/Cas9 expressing plasmids into pronucleus to generate genome edited mice. Transiently expressed gRNA/Cas9 complex digests the target locus and indels destroy gene function. Finally, we generated chimeric mice with genome-edited ES cells and analyzed gene function in mice. Our system can accelerate the reverse genetics in mice.

Free Research Field

実験動物学