ArticleNature communications2025
Cas12a Cis-cleavage mediated lateral flow assay enables multiplex and ultra-specific nucleic acid detection.
Article in Nature communications, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed.
- A field deployable duplex RPA-CRISPR/Cas12a assay for rapid and sensitive detection of African swine fever virus.Archives of virology · 2026Article
- Affinity-Activated Protein Switches for Modular and Signal-ON Detection of Small Molecules.Analytical chemistry · 2026Article
- Specific cleavage of 5' overhangs of non-target strand by Cas9 activated by target DNA binding.Nucleic acids research · 2026Article
- Structural mining and engineering of metagenome-derived Cas12a orthologs expands the CRISPR genome editing and multiplex diagnostics toolkit.Molecular therapy : the journal of the American Society of Gene Therapy · 2026Article
- crRNA Truncation MinimizesJACS Au · 2026Article
- One-Tube RPA-CRISPR/Cas12a Assays for Rapid and Visual Detection ofFoods (Basel, Switzerland) · 2026Article
- Kinetic Modeling of Target-Amplification-Free CRISPR-Cas-Based Autocatalysis Reactions.bioRxiv : the preprint server for biology · 2026Article
- Functional decoupling of crRNA enables customizable CRISPR diagnostics.Nucleic acids research · 2026Article
- Enhanced detection of HBV and HCV using Cas13a-FLAP and FGoAI platforms.Chemical science · 2026Article
- Article
- An integrated lab-in-a-tube platform for point-of-care detection of blaMikrochimica acta · 2025Article
Corrections and comments
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Authors and funding
13 authors.
Funding
Abstract
CRISPR technology holds significant promise for advancing nucleic acid assays. However, current CRISPR diagnostic techniques, reliant on indiscriminate trans-cleavage mechanisms, face challenges in developing multiplex detection formats. Moreover, chaotic trans-cleavage activity often results from mismatched targets, leading to specificity issues. To address these limitations, here we exploit a double-key recognition mechanism based on CRISPR-Cas12a cis-cleavage and invasive hybridization identification of released sticky-end DNA products. By integrating multiplexed nucleic acid amplification, the double-key Cas12a detection mechanism, and a lateral flow detection platform, we develop a method termed Cas12a cis-cleavage mediated lateral flow assay (cc-LFA). We demonstrate that the cc-LFA exhibited superior specificity compared to three mainstream trans-cleavage-based CRISPR diagnostic techniques, achieving single-base resolution detection free from high-concentration wild-type DNA background interference. cc-LFA is also applied for highly specific detection of multiple respiratory pathogen samples and precise multiplexed detection of nine high-risk human papillomavirus (HPV) subtypes, achieving over 90% sensitivity and 100% specificity, respectively. Additionally, we present a portable device to automate nucleic acid amplification and strip detection procedures, showcasing the potential of cc-LFA for future applications in decentralized laboratory scenarios.
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