CRISPR／Cas9介导的基因组定点编辑技术

New Method of Genome Editing Derived From CRISPR/Cas9

Clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated(Cas)9系统成功被改造为第三代人工核酸内切酶,与锌指核酸内切酶(zinc finger endonuclease,ZFN)和类转录激活因子效应物核酸酶(transcription activator-likeeffector nuclease,TALEN)一样可用于各种复杂基因组的编辑.目前该技术成功应用于人类细胞、斑马鱼和小鼠以及细菌的基因组精确修饰,修饰类型包括基因定点InDel突变、基因定点敲入、两位点同时突变和小片段的缺失.由于其突变效率高、制作简单及成本低的特点,被认为是一种具有广阔应用前景的基因组定点改造分子工具.本文从CRISPR/Cas的研究历史、分类、作用机理以及基因定点修饰应用等方面进行简单介绍,希望能够为在这一领域的科研工作者提供参考.

The type II clustered regularly interspaced short palindromic repeats （CRISPR）/CRISPR-associated （Cas）9 system has been used as a gene-targeting technology as ZFN and TALEN to meditate multiple genome editing. Unlike ZFN and TALEN directly binding to the specific DNA sequence to generate a double-strands break, the engineered CRISPR/Cas9 system has been demonstrated that Cas9 nuclease was directed by short RNAs to induce site-specific cleavage in complex genome. The advantage of CRPSR/Cas9 system is easy to be constructed and low cost compared with ZFN and TALEN. So, it is considered that it will replace existing technique. Here we review the CRISPR/Cas development history, classification, mechanism, progress and application of this new genome editing technology. This review will provide a useful reference for researchers who are interested in applying this new technique in their studies.