Generation of genetically modified NC/Nga mice using the CRISPR/Cas9-mediated gene editing system

2015 Fiscal Year Final Research Report

• Project/Area Number: 26670523
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Dermatology
• Research Institution: Gifu University
• Principal Investigator: Osawa Masatake 岐阜大学, 医学(系)研究科(研究院), 教授 (10344029)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: NC/Ngaマウス / ノックアウトマウス / ES細胞 / 多能性 / モデルマウス / アトピー性皮膚炎 / ゲノム編集 / CRISPR/Cas9

Outline of Final Research Achievements

In this study, we sought to develop a platform by which genetically modified mice are readily allowed to be generated in the NC/Nga strain. To this end, we generated embryonic stem cell (ESC) lines using blastocyst embryos obtained from NC/Nga mice. Using a recently developed methodology for ESC derivation from non-permissive mouse strains, we successfully established 18 ESC lines form NC/Nga blastocysts. We assayed for their chimera formation and germeline transmission capacities by 8-cell stage embryo injection. The results showed these ESCs exhibited satisfactory levels of chimera formation capacity and germline competency at early passages. However, these capacities were rapidly lost during the initial period of culture, and none of these ESCs after 6 passages showed chimera formation regardless culture conditions we tested. Therefore, we concluded that NC/Nga ESCs are extremely unstable in culture, suggestive of importance of further optimization of ECS culture conditions.

Free Research Field

皮膚科学、発生生物学