ArticleNature biotechnology2025
A resurrected ancestor of Cas12a expands target access and substrate recognition for nucleic acid editing and detection.
Article in Nature biotechnology, 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.
- Advances in prime editing: Molecular innovations, Large-fragment engineering, and AI-driven design.Biodesign research · 2026Review
- Ancient enzymes, new biotechnology applications.Nature reviews. Molecular cell biology · 2026Article
- Molecular mechanisms and biotechnology applications of CRISPR-Cas12a.Nature reviews. Molecular cell biology · 2026Review
- Substrate scope of ancestral versus modern family-1 glycosidases.Protein science : a publication of the Protein Society · 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
- A programmable bifunctional flavoenzyme for direct amine-to-ester conversion.Science advances · 2026Article
- Development of a Single-Tube Asymmetric ERA-CRISPR/Cas12a Assay for Rapid Visual Detection ofMicroorganisms · 2026Article
- Guide DNA - not RNA - expands the CRISPR toolkit.Nature biotechnology · 2026Article
- Functional decoupling of crRNA enables customizable CRISPR diagnostics.Nucleic acids research · 2026Article
- Identification of thermotolerant non-canonical PAMs for robust one-pot CRISPR-Cas12a detection.Nature communications · 2026Article
- Benchmarking the PAM compatibility of Cas12a variants for high-throughput yeast genetic variant engineering.Applied and environmental microbiology · 2025Article
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Authors and funding
10 authors.
Funding
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Abstract
The properties of Cas12a nucleases constrict the range of accessible targets and their applications. In this study, we applied ancestral sequence reconstruction (ASR) to a set of Cas12a orthologs from hydrobacteria to reconstruct a common ancestor, ReChb, characterized by near-PAMless targeting and the recognition of diverse nucleic acid activators and collateral substrates. ReChb shares 53% sequence identity with the closest Cas12a ortholog but no longer requires a T-rich PAM and can achieve genome editing in human cells at sites inaccessible to the natural FnCas12a or the engineered and PAM-flexible enAsCas12a. Furthermore, ReChb can be triggered not only by double-stranded DNA but also by single-stranded RNA and DNA targets, leading to non-specific collateral cleavage of all three nucleic acid substrates with similar efficiencies. Finally, tertiary and quaternary structures of ReChb obtained by cryogenic electron microscopy reveal the molecular details underlying its expanded biophysical activities. Overall, ReChb expands the application space of Cas12a nucleases and underscores the potential of ASR for enhancing CRISPR technologies.
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