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

Research Project

Project/Area Number 26670523
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Dermatology
Research InstitutionGifu 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
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2014: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
KeywordsNC/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 / 疾患モデルマウス / トランスジェニックマウス
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.

Report

(3 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report

URL: 

Published: 2014-04-04   Modified: 2017-05-10  

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