CRISPR/Cas9基因组编辑技术及其在动物基因组定点修饰中的应用

摘要

CRISPR/Cas system, which uses CRISPR RNAs (crRNAs) to guide Cas nuclease to silence invading nucleic acids, is self-defense system against exogenous virus or plasmid in bacteria and archaea. Through molecular modification, the typeⅡCRISPR/Cas system has become a highly efficient site-directed genome editing technique, which is simpler than zinc-finger nucleases (ZFNs) and transcription activator like effector nucleases (TALENs) and easier to be designed and applied. In this review, we summarize the evolutionary history of CRISPR/Cas9 system, the working principle and modification process of type Ⅱ CRISPR/Cas and its application in animal genome modifica-tion. We also analyze the existing problems and improvement program of the CRISPR/Cas9 system as well as its ap-plication prospect combined with successful cases, which may provide innovative perspectives on improving animal traits and establishing animal models of human diseases.%CRISPR/Cas系统是细菌和古生菌中抵抗外源病毒或质粒入侵的获得性免疫系统，利用 CRISPR RNAs (crRNAs)引导Cas核酸酶沉默入侵的核酸。通过分子生物学改造使Ⅱ型CRISPR/Cas系统成为一种高效的基因组定点修饰技术，并且比锌指核酸酶(Zinc-finger nucleases, ZFNs)和 TALE 核酸酶(Transcription activator like effector nucleases, TALENs)结构更简单，更容易设计和应用。文章主要介绍了CRISPR/Cas9系统成为高效基因组定点修饰技术的发展历程、Ⅱ型CRISPR/Cas的工作原理和改造过程以及在动物基因组定点修饰的应用，剖析了该技术存在的问题和现有改进方案，并与成功案例相结合展望了 CRISPR/Cas9系统的应用前景，以期为动物性状改良和人类疾病动物模型的创立提供新思路。