ArticleNucleic acids research2025
Unleashing high trans-substrate cleavage kinetics of Cas12a for nucleic acid diagnostics.
Article in Nucleic acids research, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 5 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
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Who cites it
5 citing papers in PubMed.
- Molecular mechanisms and biotechnology applications of CRISPR-Cas12a.Nature reviews. Molecular cell biology · 2026Review
- Toward point-of-care and amplification-free detection of human cytomegalovirus using CRISPR-Cas12a.iScience · 2026Article
- A one-pot biplex RPA-Cas assay for sensitive detection of Mycobacterium tuberculosis from tongue swabs.Scientific reports · 2026Article
- Programmable spatiotemporal control of CRISPR-Cas12a: Engineering precision for next-generation gene editing and diagnostics.Synthetic and systems biotechnology · 2026Review
- Nuc domain electrostatics drive the trans cleavage activity of CRISPR-Cas12a.Nucleic acids research · 2026Article
Corrections and comments
- Erratum issued
Authors and funding
8 authors.
Funding
Abstract
CRISPR (clustered regularly interspaced short palindromic repeats)-based nucleic acid diagnostics enable rapid, sensitive pathogen detection. Cas12a is frequently used in these assays because target-activated trans cleavage of a reporter molecule generates an easily detectable signal. However, variable activity across assays suggests that the catalytic potential of Cas12a has been limited via unknown mechanisms. Here, we show that Cas12a trans-nuclease activity is auto-inhibited by long PAM-proximal DNA (>120 bp) following cis-cleavage of targets. Short targets (<100 bp), optimized trans cleavage substrates, and low salt buffers unleash high catalytic efficiency (≈108 M-1 s-1) and turnover (≈1 s-1) across Cas12a orthologs. Pooling multiple Cas12a ribonucleoproteins (RNPs) targeting clustered protospacers overcomes cis-cleavage auto-inhibition, further boosting sensitivity. Optimized CRISPR RNA pools enable sub-femtomolar sensitivity for target detection without any pre-amplification. This mechanistic insight and mitigation strategy broaden the application of CRISPR-Cas enzymes for nucleic acid diagnostics.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.