Advances in CRISPR-based gene editing technology and its application in nucleic acid detection

Abstract

Nucleic acid analysis is a key technique that enables accurate detection of various microorganisms. Conventional nucleic acid testing typically requires access to specialized laboratories, equipment, and trained personnel, which hinders the widespread use of on-site testing for DNA and RNA targets. However, integrating gene editing technology with traditional nucleic acid detection methods, especially isothermal amplification technology, can help overcome the limitations associated with on-site testing. This combination can accomplish precise and swift detection of nucleic acid sequences, offering a robust tool for on-site detection. The Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated proteins (CRISPR/Cas) technology, which comprises the CRISPR system and Cas effector proteins, is a powerful tool that is advancing the field of nucleic acid detection. Specifically, Cas12, Cas13, and Cas14 proteins have emerged as straightforward, effective, precise, sensitive, and cost-effective methods for in vitro nucleic acid detection because of their “collateral cleavage” characteristics. When combined with the “collateral cleavage” ability of Cas protein and isothermal amplification, CRISPR/Cas systems have great potential to advance nucleic acid detection. This article summarizes the research progress of different CRISPR/Cas systems and their applications in nucleic acid detection and future perspectives.