CRISPR-Cas系统在病原核酸检测中的研究进展
Research progress of the CRISPR-Cas system in the detecting pathogen nucleic acids
杜瑶 1高宏丹 1刘家坤 2刘孝荣 3邢志浩 3张涛 4马东礼3
作者信息
- 1. 蚌埠医学院检验医学院,安徽 蚌埠 233030;深圳市儿童医院儿科研究所,广东 深圳 518034
- 2. 中国科学院深圳先进技术研究所深圳合成生物研究所,广东 深圳 518055;深圳市海微生物科技有限公司,广东 深圳 518057
- 3. 深圳市儿童医院儿科研究所,广东 深圳 518034
- 4. 蚌埠医学院病原生物学教研室,安徽 蚌埠 233030
- 折叠
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.
Abstract
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.
关键词
CRISPR-Cas系统/Cas12和Cas13/病原体/核酸检测/生物传感Key words
CRISPR-Cas system/Cas12 and Cas13/pathogen/nucleic acid detection/biosensor引用本文复制引用
基金项目
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)
出版年
2024
NETL
NSTL
万方数据