CRISPR/Cas9介导的基因编辑技术研究进展

被引：14

作者：

李聪

曹文广

机构：

[1] 中国农业科学院北京畜牧兽医研究所

来源：

生物工程学报 | 2015年 / 11期

关键词：

基因编辑; CRISPR/Cas9系统; 单链向导RNA; 脱靶效应;

D O I：

10.13345/j.cjb.140589

中图分类号：

Q78 [基因工程（遗传工程）];

学科分类号：

071007 ; 0836 ; 090102 ;

摘要：

CRISPR/Cas9(Clustered regulatory interspaced short palindromic repeats/CRISPR-associated protein 9)是最近发现的一种新型的基因组定点编辑技术。CRISPR即成簇的、有规律的、间隔短回文重复序列,是人们在研究大肠杆菌编码的碱性磷酸酶基因时发现的,它原本是存在于细菌和古细菌基因组中含有多个短重复序列的基因位点,能够为自身提供一种特异性免疫保护机制,抵御外来病毒、质粒等遗传元件的入侵。CRISPR系统主要依赖cr RNA(CRISPR RNA)和tracr RNA(Trans-activating chimeric RNA)结合并导向Cas(CRISPR-associated system)蛋白来对外源DNA进行序列特异性降解。目前已经发现了3种类型的CRISPR/Cas系统:Ⅰ型、Ⅱ型和Ⅲ型。其中Ⅱ型系统组分较为简单,主要依赖的是Cas9核心蛋白,在RNA的介导下,Cas9蛋白能够识别靶序列进行切割造成DNA的双链断裂(Double-strand breaks,DSB)。在此基础上,人们可以对基因组的特定位点进行基因打靶、基因定点插入、基因修复等各种遗传操作。这种新的基因组定点编辑技术比类转录激活因子效应物核酸酶(Transcription activator-like effector nuclease,TALEN)和锌指核酸酶(Zinc-finger nuclease,ZFN)技术设计更加简单、更容易操作,势必会有更广泛的应用。目前,利用该技术已在多个物种的细胞和个体水平上实现了遗传操作。文中将从CRISPR/Cas9的来源、结构、作用机理方面介绍其研究进展,为开展这一领域的研究工作提供参考。

引用

页码：1531 / 1542

页数：12

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