Progress in in Vitro CRISPR/Cas-based Nucleic Acid Detection
As the most advanced detection method,the sequence-based nucleic acid detection is widely used in genotyping,the diagnosis of pathogens and genetic diseases,and environmental monitoring.The CRISPR/Cas system can accurately and efficiently identify and cleave target DNA/RNA under the guidance of guide RNA,and thus is adopted to develop the new generation of in vitro nucleic acid detection with a high sensitivity and specificity.Along with the advances in multiple CRISPR/Cas systems,more and more new CRISPR/Cas-based techniques and platforms have been established for pathogen nucleic acid detection and early disease diagnosis.This article reviews the action mechanism of CRISPR/Cas9,12,13,14 systems in nucleic acid detection,and the novel signal sensing technologies of CRISPR/Cas-based nucleic acid detection,including the high-throughput and multi-channel assays,the enrichment and amplification of target nucleic acids,and the methods of signal output.It further illustrates the new application of CRISPR/Cas nucleic acid test in monitoring endogenous diseases,virus,bacteria,and parasites,discusses its advantages,disadvantages,and challenges,and outlines its future research directions,providing the reference and basis for the translation of CRISPR/Cas-based nucleic acid detection.
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