Synthetic Biology Journal ›› 2024, Vol. 5 ›› Issue (1): 202-216.DOI: 10.12211/2096-8280.2022-068

• Invited Review • Previous Articles    

Research progress of the CRISPR-Cas system in the detecting pathogen nucleic acids

Yao DU1,2, Hongdan GAO1,2, Jiakun LIU3,4, Xiaorong LIU2, Zhihao XING2, Tao ZHANG5, Dongli MA2   

  1. 1.School of Laboratory Medicine,Bengbu Medical College,Bengbu 233030,Anhui,China
    2.Pediatric Research Institute,Shenzhen Children’s Hospital,Shenzhen 518034,Guangdong,China
    3.Shenzhen Institute of Synthetic Biology,Shenzhen Institute of Advanced Technology,Chinese Academy of Sciences,Shenzhen 518055,Guangdong,China
    4.Shenzhen Hapmic Biotechnology Company limited,Shenzhen 518057,Guangdong,China
    5.Department of Pathogenic Biology,Bengbu Medical College,Bengbu 233030,Anhui,China
  • Received:2022-11-26 Revised:2023-02-15 Online:2024-03-20 Published:2024-02-29
  • Contact: Dongli MA

CRISPR-Cas系统在病原核酸检测中的研究进展

杜瑶1,2, 高宏丹1,2, 刘家坤3,4, 刘孝荣2, 邢志浩2, 张涛5, 马东礼2   

  1. 1.蚌埠医学院检验医学院,安徽 蚌埠 233030
    2.深圳市儿童医院儿科研究所,广东 深圳 518034
    3.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
    4.深圳市海微生物科技有限公司,广东 深圳 518057
    5.蚌埠医学院病原生物学教研室,安徽 蚌埠 233030
  • 通讯作者: 马东礼
  • 作者简介:杜瑶(1999—),女,硕士研究生。研究方向为病原生物学及分子生物学。 E-mail:dy852578@163.com
    马东礼(1967—),男,硕士生导师,主任技师。研究方向为病原微生物致病机制及基因精准诊断。 E-mail:madl1234@126.com
  • 基金资助:
    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

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.

Key words: CRISPR-Cas system, Cas12 and Cas13, pathogen, nucleic acid detection, biosensor

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。

关键词: CRISPR-Cas系统, Cas12和Cas13, 病原体, 核酸检测, 生物传感

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