基于CRISPR/Cas9技术构建点突变小鼠概述

Overview of the Generation of Mice Carrying Point Mutation Based on CRISPR/Cas9 Technology

2013年CRISPR(Clustered Regularly Interspaced Short Palindromic Repeats)/Cas(CRISPR-associated)系统证实能够对人的细胞和其他真核细胞进行基因组编辑,现已广泛应用于生物医学领域。本文对CRISPR在点突变小鼠构建中的应用进行概述,为单向导RNA(single guide RNA,sgRNA)和修复模板单链寡聚核苷酸(single-strand oligonucleotide,ss ODN)的设计及体外转录、受精卵显微注射、子代鼠的鉴定等提供理论参考。

CRISPR(Clustered Regularly Interspaced Short Palindromic Repeats)/Cas(CRISPR-associated)system has been widely used in biomedicine since it was first demonstrated to be able to perform genome editing in human cells and other eukaryotic cells in 2013.In this paper,the applications of CRISPR/Cas9 in the generation of mice carrying point mutation will be reviewed,providing theoretical guidance for the design and in vitro transcription of single guide RNA(sgRNA)and single strand oligonucleotide(ss ODN),for microinjection of zygotes and for genotyping of founder and progeny mice.