ArticleMolecular cell2024
Cas12a domain flexibility guides R-loop formation and forces RuvC resetting.
Article in Molecular cell, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 43 papers.
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Who cites it
43 citing papers in PubMed.
- Size and sequence variation of the Lid of the Cas12a nuclease domain results in a nickase phenotype.Nucleic acids research · 2026Article
- Architecture of a DNA-guided Cas12a.Nature structural & molecular biology · 2026Article
- Genome Editing in Solanaceae: Harnessing CRISPR-Cas Technology for Precision Crop Improvement.Plants (Basel, Switzerland) · 2026Review
- Stepwise DNA-unwinding gates TnpB genome-editing activity.Molecular cell · 2026Article
- DNA-bound avian R2 non-LTR retrotransposon protein recruits a second R2 protein for genome-protective second-strand nicking.Proceedings of the National Academy of Sciences of the United States of America · 2026Article
- Molecular mechanisms and biotechnology applications of CRISPR-Cas12a.Nature reviews. Molecular cell biology · 2026Review
- Indiscriminate Trans-Cleavage Activity of CRISPR/SuCas12a2 Enables Sensitive Detection of SARS-CoV-2.ACS sensors · 2026Article
- Helicity of the bridge helix of Cas12a regulates on-target DNA cleavage efficiency and off-target cleavage propensity.Biochemical and biophysical research communications · 2026Article
- Systematic Position Mapping of Split CRISPR-Cas12a Activators Enables Highly Sensitive miRNA Detection and Cancer Cell Stratification.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Rho (bioRxiv : the preprint server for biology · 2026Article
- Simultaneous Detection of Human Norovirus GI, GII and Hepatitis A Virus Using CRISPR-Cas12a-Based RT-RPA and Lateral Flow Strip Method.Food and environmental virology · 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
- CRISPR diagnostics: from trans-nuclease activity to cancer diagnosis.Cell & bioscience · 2026Review
- Target RNA-triggered CRISPR-Cas12a2 preferentially cleaves collateral DNA over RNA.Nucleic acids research · 2026Article
- TopCas: Topology-Gated Cas12a via DNA-RNA Chimeric Circular crRNA for Amplification-Free Nucleic Acid Detection and Conditional Gene Editing.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Target RNA-triggered CRISPR-Cas12a2 Preferentially Cleaves Collateral DNA over RNA.bioRxiv : the preprint server for biology · 2026Article
- DNA-guided CRISPR-Cas12a effectors for programmable RNA recognition and cleavage.Nature biotechnology · 2026Article
- Mechanistic basis for improved activity of Engineered AsCas12a.Communications biology · 2026Article
- Bridge helix of Cas12a is an allosteric regulator of R-loop formation and RuvC activation.Nature communications · 2026Article
- Identification of thermotolerant non-canonical PAMs for robust one-pot CRISPR-Cas12a detection.Nature communications · 2026Article
Corrections and comments
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Authors and funding
8 authors.
Funding
Abstract
The specific nature of CRISPR-Cas12a makes it a desirable RNA-guided endonuclease for biotechnology and therapeutic applications. To understand how R-loop formation within the compact Cas12a enables target recognition and nuclease activation, we used cryo-electron microscopy to capture wild-type Acidaminococcus sp. Cas12a R-loop intermediates and DNA delivery into the RuvC active site. Stages of Cas12a R-loop formation-starting from a 5-bp seed-are marked by distinct REC domain arrangements. Dramatic domain flexibility limits contacts until nearly complete R-loop formation, when the non-target strand is pulled across the RuvC nuclease and coordinated domain docking promotes efficient cleavage. Next, substantial domain movements enable target strand repositioning into the RuvC active site. Between cleavage events, the RuvC lid conformationally resets to occlude the active site, requiring re-activation. These snapshots build a structural model depicting Cas12a DNA targeting that rationalizes observed specificity and highlights mechanistic comparisons to other class 2 effectors.
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