CRISPR-associated “genetic scissors” for multiplexing analysis

CRISPR/Cas technology has greatly pushed the development of genome editing and bioanalytical sensors after its unique endonuclease activity was reported. Especially, the multiplexing sensing abilities are utilized for developing advanced disease diagnostic platforms for coping with major health emergencies, for example SARS-CoV-2 and its viral nucleic acid variants. Various disease biomarkers like nucleic acids, proteins, and bacteria can be sensitively detected. In the aspect of small molecule inspections, the CRISPR/Cas systems can also be applied to fulfill the demands. To the best of our knowledge, there is no critical review that focuses on different designs with multiplexing capabilities of the CRISPR/Cas system. Therefore, the focus of this review is on advances in CRISPR-based biosensors and multiplexing detecting platforms in recent years, with the reader being referred to earlier classical research regarding exploiting the CRISPR/Cas system. The mechanism of recognition processes, the new finding of Cas effectors, the detection of various target molecules, the multiplexing sensing strategies, and future perspectives are discussed in detail. The recent development of mass spectrometry-based strategies may open a new avenue for multiplexing CRISPR/Cas systems.