ArticleNucleic acids research2022
Improved genome editing by an engineered CRISPR-Cas12a.
Article in Nucleic acids research, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 51 papers.
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Who cites it
51 citing papers in PubMed, 98 citations in OpenAlex.
- CRISPR application in hematological disorders: from bench to bedside.Blood advances · 2026Review
- High glucose-induced mitochondrial fission promotes Müller cell activation via suppression of the Hippo pathway.Molecular and cellular biochemistry · 2026Article
- Molecular mechanisms and biotechnology applications of CRISPR-Cas12a.Nature reviews. Molecular cell biology · 2026Review
- Helicity of the bridge helix of Cas12a regulates on-target DNA cleavage efficiency and off-target cleavage propensity.Biochemical and biophysical research communications · 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
- Corydaline overcomes doxorubicin resistance in oral cancer by activating the p53-Bax-caspase axis and inhibiting MMP-driven invasion: an in vitro, in vivo, and in silico study.In vitro cellular & developmental biology. Animal · 2026Article
- Development of a multifunctional nucleic acid response platform utilizing the Cas12a and Cas13a integrated targeting system.Nucleic acids research · 2026Article
- Article
- Comparative characterization of Cas12f orthologs reveals mechanistic features underlying enhanced genome editing efficiency.Nature structural & molecular biology · 2026Article
- Engineered CRISPR-Cas13a system with enhanced target RNA cleavage activity and reduced collateral activity for therapeutic applications.Molecular therapy. Nucleic acids · 2026Article
- The emerging impact of CRISPR and gene editing on global crop improvement.Transgenic research · 2026Review
- Bridge helix of Cas12a is an allosteric regulator of R-loop formation and RuvC activation.Nature communications · 2026Article
- Bacterial defense systems: Mechanisms, homology to eukaryotic immune systems, and applications.Zoological research · 2026Review
- CRISPR-driven strategies to disrupt methicillin-resistantFrontiers in cellular and infection microbiology · 2026Review
- Multilayer regulation of CRISPR systems: integrating anti-CRISPR proteins, CRISPRi/a, and quorum sensing networks.Frontiers in cellular and infection microbiology · 2026Review
- A buffer-tuning strategy to profile domain-specific activity of chimeric I-TevI/CRISPR gene editors in vitro.Scientific reports · 2025Article
- Efficient and multiplexed somatic genome editing with Cas12a mice.Nature biomedical engineering · 2025Article
- Tailoring Cas12a functionality with a user-friendly and versatile crRNA variant toolbox.Nature communications · 2025Article
- A resurrected ancestor of Cas12a expands target access and substrate recognition for nucleic acid editing and detection.Nature biotechnology · 2025Article
- Lung and liver editing by lipid nanoparticle delivery of a stable CRISPR-Cas9 ribonucleoprotein.Nature biotechnology · 2025Article
Corrections and comments
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Authors and funding
10 authors at 3 institutions in 1 country.
Funding
Abstract
CRISPR-Cas12a is an RNA-guided, programmable genome editing enzyme found within bacterial adaptive immune pathways. Unlike CRISPR-Cas9, Cas12a uses only a single catalytic site to both cleave target double-stranded DNA (dsDNA) (cis-activity) and indiscriminately degrade single-stranded DNA (ssDNA) (trans-activity). To investigate how the relative potency of cis- versus trans-DNase activity affects Cas12a-mediated genome editing, we first used structure-guided engineering to generate variants of Lachnospiraceae bacterium Cas12a that selectively disrupt trans-activity. The resulting engineered mutant with the biggest differential between cis- and trans-DNase activity in vitro showed minimal genome editing activity in human cells, motivating a second set of experiments using directed evolution to generate additional mutants with robust genome editing activity. Notably, these engineered and evolved mutants had enhanced ability to induce homology-directed repair (HDR) editing by 2-18-fold compared to wild-type Cas12a when using HDR donors containing mismatches with crRNA at the PAM-distal region. Finally, a site-specific reversion mutation produced improved Cas12a (iCas12a) variants with superior genome editing efficiency at genomic sites that are difficult to edit using wild-type Cas12a. This strategy establishes a pipeline for creating improved genome editing tools by combining structural insights with randomization and selection. The available structures of other CRISPR-Cas enzymes will enable this strategy to be applied to improve the efficacy of other genome-editing proteins.
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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.