Simultaneous generation and germline transmission of multiple gene mutations in rat using CRISPR-Cas systems

摘要

CRISPRs are clustered, regularly interspaced, short palindromic repeats present in many bacteria and archaea genomes. Proteins encoded by CRISPR-associated (Cas) genes serve as guardians of the genome, which target foreign DNA at specific sites by means of small CRISPR RNA (crRNA)-guided DNA recognition and degradation1"4. Recently, several groups described how CRISPR-Cas systems efficiently create site-specific gene modifications in whole organisms such as Streptococcus pneumoniae, Escherichia coli,Danio rerio (zebrafish) and mice, suggesting its potential application in the production of genetically engineered organisms5-7, although germline transmission of the mutations remains to be shown. Here, we report the use of CRISPR-Cas systems to generate multiple gene mutations in rats in a germline-competent manner.