摘要
CRISPR-Cas系统作为原核生物获得性免疫系统,由簇状规则间隔短回文重复序列(clustered regularly interspaced short palindromic repeats,CRISPR)和CRISPR相关蛋白(CRISPR-associated proteins,Cas)构成,因其识别和切割特定DNA或RNA序列,而成为分子诊断领域研究的热点。研究人员利用Cas蛋白(Cas12、Cas13、Cas14、Cas3等)结合信号放大和转化技术(荧光法、电位法、比色法、侧向流动技术等),开发了许多高灵敏度、高特异性、低成本的诊断平台,为病原核酸检测提供了新途径。本文介绍了CRISPR-Cas系统的生物学机制及分类,总结现有的基于Cas蛋白反式切割活性开发的病原核酸检测技术,描述其特点、功能和应用场景,并对该系统的未来应用前景进行展望,期望CRISPR-Cas系统成为包括核酸检测在内的多靶标的理想检测平台。
The CRISPR-Cas system consists of clustered regularly interspaced short palindromic repeats(CRISPR)and CRISPR-associated(Cas)proteins,which has become the focus of molecular diagnosis because it recognizes and cleaves specific DNA or RNA sequences.Using Cas proteins(Cas12,Cas13,Cas14,Cas3,etc.)combined with signal amplification and transformation techniques(fluorescence,potentiometric,colorimetric,lateral flow assay,etc.),researchers have developed many diagnostic platforms with high sensitivity,good specificity,and low cost,which provide a new tool for detecting pathogen nucleic acids.This review presents the biological mechanism and classification of the CRISPR-Cas system,and also summarizes existing technologies for detecting pathogenic nucleic acids based on the trans-cleavage activity of Cas proteins,commenting their properties,functions and application scenarios,with future applications prospected based on the functional characteristics of the CRISPR-Cas system,which is expected to become an ideal detection platform for other multiple targets.
作者
杜瑶
高宏丹
刘家坤
刘孝荣
邢志浩
张涛
马东礼
DU Yao;GAO Hongdan;LIU Jiakun;LIU Xiaorong;XING Zhihao;ZHANG Tao;MA Dongli(School of Laboratory Medicine,Bengbu Medical College,Bengbu 233030,Anhui,China;Pediatric Research Institute,Shenzhen Children’s Hospital,Shenzhen 518034,Guangdong,China;Shenzhen Institute of Synthetic Biology,Shenzhen Institute of Advanced Technology,Chinese Academy of Sciences,Shenzhen 518055,Guangdong,China;Shenzhen Hapmic Biotechnology Company limited,Shenzhen 518057,Guangdong,China;Department of Pathogenic Biology,Bengbu Medical College,Bengbu 233030,Anhui,China)
出处
《合成生物学》
CSCD
北大核心
2024年第1期202-216,共15页
Synthetic Biology Journal
基金
广东省高水平医院项目(SEY-GSP-YXPT-A02)
深圳市发展和改革委员会赠款(2019)986。
