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

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.