新一代精准基因编辑工具CRISPR/Cas9的技术优势与应用局限

The Advantages and Limitations of CRISPR/Cas9-based Gene Editing Technology

CRISPR/Cas9(Clustered regularly interspaced short palindromic repeats/Cas9)是继锌指核酸酶(ZFNs)和类转录激活因子效应物核酸酶(TALENs)基因编辑技术之后的第三代基因编辑技术。CRISPR/Cas9在细菌和古生菌中广泛存在,是细菌在长期进化过程中形成的一种"适应性免疫防御",能够针对噬菌体感染、质粒接合和转化所造成的外源导入基因特异性识别、降解入侵的外源DNA,CRISPR/Cas9通过一段20 bp的短RNA来识别打靶位点的精准编辑技术。CRISPR/Cas9具有设计操作简便、编辑高效和通用性广等优点,是新一代具有革命意义的精准基因编辑技术。从CRISPR/Cas9的发现、作用机理、基因编辑以及应用局限等方面进行归纳总结,旨为理解其工作原理和精准基因编辑技术应用提供参考。

CRISPR/Cas9 is becoming a hot spot in genome editing,and it provides a more efficient way for gene targeting than zinc finger endonuclease（ZFN）and transcription activator-like effector nuclease（TALEN）. CRISPR/Cas9 system exists widely in bacteria andarchaea,carrying out the adaptive immune responses specifically recognizing the invaded genetic elements from phage infection,plasmidconjugation and transformation as well as and degrading exogenous DNA. CRISPR/Cas9 is a precise editing technology by which the target sitesare recognized by a segment of 20 bp small RNA. CRISPR/Cas9 has the advantages of simple design and operation,high efficiency,and broadgenerality,thus it is a novel and revolutionary precise gene editing technology. This review induced and summarized the aspects of the discoveryof CRISPR/Cas,mechanisms,gene editing,and application limitations,aiming at providing references for understanding its mechanisms and applying precise gene editing technology.