基于CRISPR/Cas13的RNA编辑系统及其在核酸检测中的应用

RNA Editing System Based on CRISPR/Cas13 and Its Application in Nucleic Acid Detection

核酸检测技术在农业、医药、环境等多个领域均有广泛应用,传统的核酸检测技术存在设备昂贵、操作复杂和灵敏度低等诸多局限性。成簇规律间隔短回文重复序列及其相关基因13(CRISPR/Cas13)是一种只靶向RNA的新型CRISPR系统,能在CRISPR RNA(crRNA)引导下特异切割单链靶RNA,并有非特异性的“附带切割”能力。基于CRISPR/Cas13的核酸检测技术具有灵敏度高、特异性强、操作方便、成本低廉等优点,具有广阔的应用前景。阐述了CRISPR/Cas13系统的类别归属及其亚型,组分特征及其分子作用机制,介绍了基于CRISPR/Cas13系统的特异性高灵敏度酶报告解锁(SHERLOCK)技术的产生和发展,综述了CRISPR/Cas13系统在病原体检测、耐药突变检测、物种识别和转基因鉴定方面的应用,同时指出了目前基于CRISPR/Cas13核酸检测技术存在的问题,并对未来应用进行了展望。

Nucleic acid detection technology is widely used in agriculture,medicine,environment and other fields.The traditional nucleic acid detection technology has many limitations such as expensive equipment,complex operation and low sensitivity.Clustered regularly interspaced short palindromic repeats/CRISPR-associated gene 13(CRISPR/Cas13)is a novel CRISPR system that only targets RNA.It can specifically cleave single strand target RNA under the guidance of crRNA,and has the ability of nonspecific“collateral cleavage”.The nucleic acid detection technology based on CRISPR/Cas13 has the advantages of high sensitivity,strong specificity,convenient operation and low cost,which has broad application prospects.The classification and subtypes of CRISPR/Cas13 system and its component characteristics and molecular mechanism are reviewed,the generation and development of specific high-sensitivity enzymatic reporter unlocking(SHERLOCK)technology based on CRISPR/cas13 system is introduced,and its application in pathogen detection,drug resistance mutation detection,species identification and transgenic identification is summarized.At the same time,the existing problems based on CRISPR/Cas13 nucleic acid detection technology are pointed out,and the future application is prospected.