基于CRISPR/Cas系统的病原微生物检测体系构建研究进展

Research progress in construction of pathogenic microorganism detection system based on CRISPR/Cas system

摘要:

规律成簇间隔短回文重复序列及其相关蛋白(clustered regularly interspaced short palindromic repeats/CRISPR-associated protein,CRISPR/Cas)自发现以来,其应用范围在不断拓宽。除在基因编辑方面出色的应用能力外,近年来Cas12、Cas13等蛋白反式切割活性的发现,使其在病原微生物快速检测方面也显现出巨大的应用潜力。基于CRISPR/Cas系统建立的病原微生物检测技术具有灵敏度高、特异性强、操作简单等特点,通过设计不同靶向的引导RNA能够对不同的目标进行快速检测,是一种极具应用潜力的检测技术。本文对基于CRISPR/Cas开发的部分代表性病原微生物检测技术类型进行总结。由于引导RNA在CRISPR/Cas系统中具有重要作用,但目前鲜见相关总结文章,本文对部分Cas蛋白引导RNA特点以及特异性靶标的获取也进行了重点阐述,以期为开发基于CRISPR/Cas系统新型病原微生物检测技术提供参考和依据。

Abstract:

The clustered regularly interspaced short palindromic repeats/CRISPR-associated protein (CRISPR/Cas) has been applied in increasing fields since its discovery.Beyond gene editing,certain Cas proteins such as Cas12 and Cas13 have shown great application potential in rapid detection of pathogenic microorganisms since the discovery of their trans-cleavage activity.The CRISPR/Cas-based pathogenic microorganism detection technology is characterized by high sensitivity,high specificity,and simple operation.The presence/absence of targeted microorganisms can be quickly determined with help of the specific guide RNAs designed by bioinformatic analysis.This review introduces the representative CRISPR/Cas-based pathogenic microorganism detection technologies.Further,we expound the characteristics of guide RNAs required by different CRISPR/Cas complexes and provide guidance on how to design the guide RNAs used in the CRISPR/Cas-based pathogenic microorganism detection systems,given the importance of guide RNA in the CRISPR/Cas-based detection system and the lack of comprehensive summarization in this topic.This review aims to contribute to the development of CRISPR/Cas-based pathogenic microorganism detection system.