biostudies-literatureMoon JU.S. Department of Health & Human Services | NIH | National Institute of Allergy and Infectious DiseasesNIAID NIH HHSU.S. Department of Health & Human Services | NIH | National Cancer InstituteNCI NIH HHS7504https://www.ebi.ac.uk/biostudies/studies/S-EPMC10657364biostudies-literatureUnknown14(1)Nucleic acid detection powered by CRISPR technology provides a rapid, sensitive, and deployable approach to molecular diagnostics. While exciting, there remain challenges limiting its practical applications, such as the need for pre-amplification and the lack of quantitative ability. Here, we develop an asymmetric CRISPR assay for cascade signal amplification detection of nucleic acids by leveraging the asymmetric trans-cleavage behavior of competitive crRNA. We discover that the competitive reaction between a full-sized crRNA and split crRNA for CRISPR-Cas12a can induce cascade signal amplification, significantly improving the target detection signal. In addition, we find that CRISPR-Cas12a can recognize fragmented RNA/DNA targets, enabling direct RNA detection by Cas12a. Based on these fNature communicationsAsymmetric CRISPR enabling cascade signal amplification for nucleic acid detection by competitive crRNA.PMC10657364U01CA269147U01 CA269147U01AI148306U01 AI148306Liu CMoon JfalseAsymmetric CRISPR enabling cascade signal amplification for nucleic acid detection by competitive crRNA.Nucleic acid detection powered by CRISPR technology provides a rapid, sensitive, and deployable approach to molecular diagnostics. While exciting, there remain challenges limiting its practical applications, such as the need for pre-amplification and the lack of quantitative ability. Here, we develop an asymmetric CRISPR assay for cascade signal amplification detection of nucleic acids by leveraging the asymmetric trans-cleavage behavior of competitive crRNA. We discover that the competitive reaction between a full-sized crRNA and split crRNA for CRISPR-Cas12a can induce cascade signal amplification, significantly improving the target detection signal. In addition, we find that CRISPR-Cas12a can recognize fragmented RNA/DNA targets, enabling direct RNA detection by Cas12a. Based on these f2023-01-01T00:00:00Z2023 Nov2025-04-19T06:36:05.302Z2025-04-19T06:36:05.302ZS-EPMC106573643798040410.1038/s41467-023-43389-7