ZFNs、TALENs和CRISPR-Cas基因组靶向编辑技术及其在植物中的应用  

ZFNs,TALENs and CRISPR-Cas Targeted Genome Editing Technologies and Their Applications in Plants

作　　者：廖鹏飞[1] 聂旺[1] 余雅心 童普国 李绍波[1] 朱友林[1] 

机构地区：[1]南昌大学生命科学学院,江西省分子生物学与基因工程重点实验室,南昌330031

出　　处：《基因组学与应用生物学》2016年第2期442-451,共10页Genomics and Applied Biology

基　　金：国家自然科学基金(31260313);江西省自然科学基金(20132BAB204012)共同资助

摘　　要：锌指核酸酶(zinc-finger nucleases,ZFNs)、转录激活子样效应因子核酸酶(transcription activator-like effectors nucleases,TALENs)和RNA介导的CRISPR-Cas系统是当今三种主要的基因组靶向编辑技术,ZFNs和TALENs由特异性的DNA结合蛋白融合一个非特异性的核酸酶FokⅠ组成,DNA结合蛋白特异识别并结合靶DNA序列,然后在FokⅠ的作用下引起靶位点的DNA双链断裂(double strand breaks,DSBs);而CRISPR-Cas系统,则是由小分子向导RNA通过碱基互补配对与靶基因组序列结合,而引导Cas核酸酶切割靶位点而引起DSBs。DSBs通过真核细胞DNA修复机制(非同源末端连接和同源重组)进行修复,从而实现基因组靶向编辑。本综述着重介绍这三种技术的基本结构及其基因组靶向编辑的原理和特点,对三种技术在植物中的应用进展进行介绍并对其进一步的发展提出了展望。ZFNs （zinc-finger nucleases）, TALENs （transcription activator-like effector nucleases） and the RNA- guided CRISPR-Cas nuclease system are three main technologies for targeted genome editing. ZFNs and TALENs are composed of a specific DNA-binding protein and a non-specific Fok I nuclease domain, the DNA-binding protein bind specifically to the target DNA, and then, double strand breaks （DSBs） are introduced at the target DNA site via the non-specific Fok I nuclease. In the CRISPR-Cas, on the other hand, a small guide RNA base paring with complementary the target genome sequence, leads Cas nuclease to introduce DSBs at targeted site. The precise genome editing will be accomplished when these DSBs are repaired by eukaryotic cell DNA-repair mechanism involving either non-homologous end joining or homologous recombination. This review focused on describing the structures and principles of the above three genome editing technologies, and summarizing their applications in plants and discussing future prospects.

关 键 词：基因组靶向编辑技术 ZFNs TALENs CRISPR/Cas 基因打靶 DNA双链断裂(DSBs) 

分 类 号：Q943.2[生物学—植物学]