Generation of site-specific mutations in the rat genome via CRISPR/Cas9.
The laboratory rat is a valuable model organism for basic biological studies and drug development. However, due to the lack of genetic tools for site-specific genetic modification in the rat genome, more and more researchers chose the mouse as their favored mammalian models due to the sophisticated embryonic stem cell-based gene-targeting techniques available. Recently, engineered nucleases, including zinc finger nucleases, transcription activator-like effector nucleases, and CRISPR/Cas9 systems, have been adapted to generate knockout rats efficiently. The purpose of this section is to provide detailed procedures for the generation of site-specific mutations in the rat genome through injection of Cas9/sgRNA into one-cell embryos.
Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China.
Yuting
Y
Guan
Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China.
Yanjiao
Y
Shao
Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China. Electronic address: dlli@bio.ecnu.edu.cn.
Dali
D
Li
Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China. Electronic address: myliu@bio.ecnu.edu.cn.
Mingyao
M
Liu
D000818:Animals; D064130:CRISPR-Associated Proteins / Q000235:genetics; D064113:CRISPR-Cas Systems; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004622:Embryo, Mammalian / Q000166:cytology* / Q000378:metabolism; D004720:Endonucleases / Q000235:genetics; D005260:Female; D018390:Gene Targeting / Q000379:methods*; D016678:Genome; D016296:Mutagenesis; D009154:Mutation; D051381:Rats
D064130:CRISPR-Associated Proteins; D004720:Endonucleases
Gene editing; Knockin; Knockout; Mutation; One-cell embryo; Rat genome