基于不同Cas9突变体的CRISPR/Cas9在医学检验新方法领域的发展与应用  

Development and Application of Different CRISPR/Cas9 Mutants in New Methods of Medical Examination

作　　者：陈卓颖 戴枫 段晓雷 CHEN Zhuoying;DAI Feng;DUAN Xiaolei(Department of Laboratory Medicine,the Affiliated Hospital of Zunyi Medical College,Zunyi 563003,China;Department of Clinical Biochemical Examination,School of Laboratory Medicine,Zunyi Medical University,Zunyi 563006,China)

机构地区：[1]遵义医科大学附属医院医学检验科,贵州遵义563003 [2]遵义医科大学检验医学院临床生物化学检验教研室,贵州遵义563006

出　　处：《医学综述》2022年第24期4939-4943,共5页Medical Recapitulate

基　　金：国家自然科学基金(82160409);重庆市基础研究与前沿探索项目(重庆市自然科学基金)(cstc2018jcyjAX0201);贵州省科技计划项目(黔科合基础〔2018〕1196)。

摘　　要：CRISPR/Cas9核酸编辑系统具有组成简易、靶向性强、切割效率高等特点,其在医学检验新方法领域的研究与应用日益广泛。具有不同结构域Cas9核酸酶的CRISPR/Cas9的活性差异明显,并基于此研究出不同功能的Cas9突变体蛋白,包括同时切割双链DNA的野生型、只切割目标链的RuvC突变型、只切割非目标链的HNH突变型、只结合不切割的双突变型。近年来,基于上述CRISPR/Cas9体系发展出众多高特异度、高灵敏度、简便快捷检测靶标分子的新方法,且其在肿瘤、病原体早期诊断技术研发中具有巨大的应用潜力。但在未来医学检验新方法研究中,克服脱靶效应是CRISPR/Cas9需要解决的主要问题。CRISPR/Cas9 genome editing system has been widely studied and used in new methods of laboratory medicine for its simple composition,strong targeting and high cutting efficiency.There is significant difference in the activity of CRISPR/Cas9 of Cas9 nucleasewith different domains,based on which Cas9 mutant proteins with different functions are studied,including the wild type with simultaneous cleavage activity of double stranded DNA,the RuvC domain mutant type with cleavage of only the target DNA chain,the HNH domain mutant type with cleavage of only the non-target DNA chain and the double domain mutant type with only binding but not cleavage activity.In recent years,based on these different CRISPR/Cas9 systems,researchers have developed many new methods for the detection of target molecules with high specificity,high sensitivity and convenience,which have great potentials in the development of early diagnostic technologies for tumor and pathogens.However,how to solve the off-target effect of CRISPR/Cas9 is the major issue need to be addressed in new methods of laboratory medicine in future.

关 键 词：CRISPR/Cas9系统 突变体蛋白 结构域 脱靶效应 

分 类 号：R541.4[医药卫生—心血管疾病]