ArticleNucleic acids research2023
Unidirectional trans-cleaving behavior of CRISPR-Cas12a unlocks for an ultrasensitive assay using hybrid DNA reporters containing a 3' toehold.
Article in Nucleic acids research, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Attomolar Detection of HIV-1 With Label-Free RCA-rCRISPR on Smartphone.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Molecular mechanisms and biotechnology applications of CRISPR-Cas12a.Nature reviews. Molecular cell biology · 2026Review
- RAPID: evaluation of Cas12a protospacer nicking and chimeric reporters for PAM-independent RNA and DNA diagnostics.Nucleic acids research · 2026Article
- Target RNA-triggered CRISPR-Cas12a2 preferentially cleaves collateral DNA over RNA.Nucleic acids research · 2026Article
- Target RNA-triggered CRISPR-Cas12a2 Preferentially Cleaves Collateral DNA over RNA.bioRxiv : the preprint server for biology · 2026Article
- Kinetic Modeling of Target-Amplification-Free CRISPR-Cas-Based Autocatalysis Reactions.bioRxiv : the preprint server for biology · 2026Article
- Programmable hooded DNA switches for conditional control of CRISPR/Cas12a in multiplexed biosensing.Journal of nanobiotechnology · 2026Article
- Identification of thermotolerant non-canonical PAMs for robust one-pot CRISPR-Cas12a detection.Nature communications · 2026Article
- Amplification-Free Dual-Blocking Autocatalytic CRISPR-Cascade for Atto-Molar DNA Detection with Low Nonspecific Signal.bioRxiv : the preprint server for biology · 2025Article
- One-Pot Isothermal Nucleic Acid Amplification Assisted CRISPR/Cas Detection Technology: Challenges, Strategies, and Perspectives.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Review
- Cas12a Cis-cleavage mediated lateral flow assay enables multiplex and ultra-specific nucleic acid detection.Nature communications · 2025Article
- Enhancement of CRISPR-Cas12a system through universal circular RNA design.Cell reports methods · 2025Article
- Expanding Cas12a Activity Control with an RNA G-Quadruplex at the 5' end of CRISPR RNA.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Ratiometric nonfluorescent CRISPR assay utilizing Cas12a-induced plasmid supercoil relaxation.Communications chemistry · 2024Article
- An autocatalytic CRISPR-Cas amplification effect propelled by the LNA-modified split activators for DNA sensing.Nucleic acids research · 2024Article
Corrections and comments
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Authors and funding
5 authors.
Funding
Abstract
CRISPR-Cas12a can induce nonspecific trans-cleavage of dsDNA substrate, including long and stable λ DNA. However, the mechanism behind this is still largely undetermined. In this study, we observed that while trans-activated Cas12a didn't cleave blunt-end dsDNA within a short reaction time, it could degrade dsDNA reporters with a short overhang. More interestingly, we discovered that the location of the overhang also affected the susceptibility of dsDNA substrate to trans-activated Cas12a. Cas12a trans-cleaved 3' overhang dsDNA substrates at least 3 times faster than 5' overhang substrates. We attributed this unique preference of overhang location to the directional trans-cleavage behavior of Cas12a, which may be governed by RuvC and Nuc domains. Utilizing this new finding, we designed a new hybrid DNA reporter as nonoptical substrate for the CRISPR-Cas12a detection platform, which sensitively detected ssDNA targets at sub picomolar level. This study not only unfolded new insight into the trans-cleavage behavior of Cas12a but also demonstrated a sensitive CRISPR-Cas12a assay by using a hybrid dsDNA reporter molecule.
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