基于CRISPR/Cas技术的核酸检测研究进展

Research progress of nucleic acid detection based on CRISPR/Cas technology

摘要:

由成簇、规则间隔的短回文重复序列（Clustered regularly interspaced short palindromic repeats，CRISPR）和CRISPR相关蛋白（CRISPR-associated protein，Cas）组成的CRISPR/Cas系统是广泛存在于多数细菌和古细菌中的一种适应性免疫系统。CRISPR/Cas系统可识别并结合外源入侵的核酸分子，之后Cas蛋白的切割活性被激活，能够对入侵的核酸分子进行切割使其降解。利用CRISPR/Cas系统特异的序列识别及切割活性，将其应用于核酸检测中，为提高检测灵敏度及特异性等性能指标提供了一种新思路。本文介绍了CRISPR/Cas系统的发展、作用机制等，对多样化的Cas蛋白在核酸检测中的代表性应用研究进行总结，进一步讨论了CRISPR/Cas技术应用于核酸检测中存在的优缺点，并对未来研究进行了展望，为基于CRISPR/Cas技术的核酸检测方法在病原微生物的检测中提供参考和依据。

关键词:CRISPR/Cas;Cas蛋白;切割活性;核酸检测;病原微生物

Abstract:

The CRISPR/Cas system, an adaptive immune system widely found in most bacteria and archaea, is composed of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (Cas). The CRISPR/Cas system can recognize and bind foreign invading nucleic acid. Then the cleavage activity of Cas protein is activated that can cleave and degrade the invasive nucleic acid. The specific sequence recognition and cleavage activity of the CRISPR/Cas system is applied to nucleic acid detection that provides a new research idea to improve detection performance such as detection sensitivity and specificity. This study introduces the development of the CRISPR/Cas system, its mechanism of action, etc. In addition, we also summarize the representative applications of diversified Cas proteins in nucleic acid detection. The advantages and disadvantages of CRISPR/Cas technology in nucleic acid detection are discussed, and future research is proposed. In sum, this review aims to provide references for the detection of pathogenic microorganisms by using a nucleic acid detection method based on CRISPR/Cas technology.

Key words:CRISPR/Cas;Cas protein;cleavage activity;nucleic acid detection;pathogenic microorganism