Rapid generation of conditional knockout mice using the CRISPR-Cas9 system and electroporation for neuroscience research

被引：10

作者：

Nishizono, Hirofumi ^{[1
,2
]}

Hayano, Yuki ^{[2
]}

Nakahata, Yoshihisa ^{[2
]}

Ishigaki, Yasuhito ^{[1
]}

Yasuda, Ryohei ^{[2
]}

机构：

[1] Kanazawa Med Univ, Med Res Inst, 1-1 Daigaku, Uchinada, Ishikawa 9200293, Japan

[2] Max Planck Florida Inst Neurosci, 1 Max Planck Way, Jupiter, FL 33458 USA

来源：

MOLECULAR BRAIN | 2021年 / 14卷 / 01期

关键词：

Cre; LoxP; floxed mouse; CRISPR-Cas9; genome editing; CaMK1; GENOME;

D O I：

10.1186/s13041-021-00859-7

中图分类号：

Q189 [神经科学];

学科分类号：

071006 ;

摘要：

The Cre/LoxP-based conditional knockout technology is a powerful tool for gene function analysis that allows region- and time-specific gene manipulation. However, inserting a pair of LoxP cassettes to generate conditional knockout can be technically challenging and thus time- and resource-consuming. This study proposes an efficient, low-cost method to generate floxed mice using in vitro fertilization and the CRISPR-Cas9 system over two consecutive generations. This method allowed us to produce floxed mice targeting exons 5 and 6 of CaMK1 in a short period of 125 days, using only 16 mice. In addition, we directly edited the genome of fertilized eggs of mice with our target genetic background, C57BL/6 N, to eliminate additional backcrossing steps. We confirmed that the genome of the generated floxed mice was responsive to the Cre protein. This low-cost, time-saving method for generating conditional knockout will facilitate comprehensive, tissue-specific genome analyses.

引用

页数：5

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