Strategies and challenges of CRISPR/Cas system in detecting foodborne pathogens

Nucleic acid-based alternatives, including polymerase chain reaction, nucleic acid hybridization, and isothermal amplification techniques, satisfy the requirement of rapid and accurate detection. However, the majority of the established methods necessitate skilled operators and a well-established laboratory with expensive equipment, which constrains their usage in low-resource areas. Currently, clustered regularly interspaced short palindromic repeats/CRISPR-associated protein (CRISPR/Cas) systems, known for their accuracy in recognizing specific nucleic acid sequences, are attracting attention as biosensing platforms, particularly after the discovery of the trans-cleavage activity of several Cas proteins. This chapter presents the features of the class II Cas proteins, the fundamentals of CRISPR guide RNA design, and the CRISPR/Cas reporter molecules. This is followed by an overview of several CRISPR/Cas-based foodborne pathogens detection strategies and a discussion of the challenges of the CRISPR/Cas system.