CRISPR molecular detection techniques: Advances from single to multiple detection methods

The clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-related proteins (CRISPR/Cas) are regarded as the next generation of molecular diagnostic systems. Due to its good biocompatibility, unique cutting activity, ease of operation and design, the CRISPR/Cas system plays an important role in biosensing applications, especially the trans-cutting activity of several Cas effectors. In recent years, CRISPR/Cas, as an emerging molecular recognition component and signal transduction module, has been combined with modern analytical techniques for detecting various analytes. This paper briefly introduces the classification and working principle of CRISPR sensor-related enzymes, summarizes the latest research on the detection of various Cas effectors on single (multiple) targets, and analyzes their application examples in practical and portable sensing from the perspective of multiple detection platforms. Finally, the technical bottleneck and application prospects were summarized.