CRISPR/Cas9基因编辑原理、发展及应用

CRISPR/Cas9 Gene Editing:Principle,Development and Application

CRISPR/Cas9(Clustered regularly interspaced short palindromic repeats/Cas9)是一个操作便捷、编辑高效和通用性广的基因编辑工具,可使快速而精准地阐明基因组的结构和功能,在遗传变异和生物表型之间建立因果联系。它是由一个非特异性的Cas9核酸酶和一组可与靶基因特异性互补的CRISPR RNA(crRNA)组成的精准编辑系统。通过20 bp的crRNA和位于靶序列下游的原间隔序列邻近基序(PAM)来决定CRISPR/Cas9实现DNA切割的特异性。经过遗传工程改造后的CRISPR/Cas9已经作为一种新型的基因编辑工具被用于多种生物的基因组编辑。本文通过综述CRISPR/Cas9的历史来源、组成结构、作用机理、技术发展和应用等,旨在为其在水产方面得到广泛应用提供理论基础。

The CRISPR/Cas9(Clustered regularly interspaced short palindromic repeats/Cas9)system provides genomic editing with a wide range of tools for easy-to-use,editing efficiency,and versatility,enabling researchers to articulate the functional tissue of the genome at the system level and to establish causal connections between genetic variation and phenotype.CRISPR/Cas9 consists of a nonspecific Cas9 nuclease and a set of programmable sequence-specific CRISPR RNA(crRNA)to precise edit genome on target sites comprised of a short sequence of 20 bp.The specificity of CRISPR/Cas9-mediated DNA cutting requires the target sequence to match crRNA and the protective base sequence adjacent to the sequence(PAM)located downstream of the target sequence.Genetically engineered CRISPR/Cas9 has been used as a new gene editing tool for genome editing of many organisms.This review summarizes the history,structure,function mechanism,technological development and application of the CRISPR/Cas9 system,which provides a basic knowledge on genome edit for researchers.