ArticleBMC biology2022
Engineered Cas12a-Plus nuclease enables gene editing with enhanced activity and specificity.
Article in BMC biology, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 51 citations in OpenAlex.
- Advanced gene editing technologies for oncology mechanisms, applications, and clinical implementation.Cancer gene therapy · 2026Review
- Molecular mechanisms and biotechnology applications of CRISPR-Cas12a.Nature reviews. Molecular cell biology · 2026Review
- Evaluating high-fidelity CRISPR-Cas nucleases in nucleosomal contexts using a quantitative framework.Frontiers in genome editing · 2026Article
- Prime editing in mammals: From promise to practicalities.Molecular therapy. Nucleic acids · 2025Review
- Understanding the stability landscape of LbCas12a by deep analysis of stabilizing mutations and mutation combinations.Protein science : a publication of the Protein Society · 2025Article
- Never-germinating Arabidopsis seeds with LbCas12a-induced mutations in 6 clade A type 2C protein phosphatase genes.Plant physiology · 2025Article
- Highly parallel profiling of the activities and specificities of Cas12a variants in human cells.Nature communications · 2025Article
- Biology and applications of CRISPR-Cas12 and transposon-associated homologs.Nature biotechnology · 2024Review
- Review
- CRISPR-array-mediated imaging of non-repetitive and multiplex genomic loci in living cells.Nature methods · 2024Article
- Enhanced editing efficiency in Arabidopsis with a LbCas12a variant harboring D156R and E795L mutations.aBIOTECH · 2024Article
- Engineering of Cas12a nuclease variants with enhanced genome-editing specificity.PLoS biology · 2024Article
- High-Fidelity Identification of Single Nucleotide Polymorphism by Type V CRISPR Systems.ACS sensors · 2023Article
- Discovery of Diverse CRISPR-Cas Systems and Expansion of the Genome Engineering Toolbox.Biochemistry · 2023Review
- Unleashing the potential of CRISPR multiplexing: Harnessing Cas12 and Cas13 for precise gene modulation in eye diseases.Vision research · 2023Review
- Characterization of a thermostable Cas12a ortholog.Cell insight · 2023Article
- CRISPR applications in cancer diagnosis and treatment.Cellular & molecular biology letters · 2023Review
- Engineered circular guide RNAs boost CRISPR/Cas12a- and CRISPR/Cas13d-based DNA and RNA editing.Genome biology · 2023Article
- Profiling the impact of the promoters on CRISPR-Cas12a system in human cells.Cellular & molecular biology letters · 2023Article
- Boosting genome editing efficiency in human cells and plants with novel LbCas12a variants.Genome biology · 2023Article
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Authors and funding
15 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundThe CRISPR-Cas12a (formerly Cpf1) system is a versatile gene-editing tool with properties distinct from the broadly used Cas9 system. Features such as recognition of T-rich protospacer-adjacent motif (PAM) and generation of sticky breaks, as well as amenability for multiplex editing in a single crRNA and lower off-target nuclease activity, broaden the targeting scope of available tools and enable more accurate genome editing. However, the widespread use of the nuclease for gene editing, especially in clinical applications, is hindered by insufficient activity and specificity despite previous efforts to improve the system. Currently reported Cas12a variants achieve high activity with a compromise of specificity. Here, we used structure-guided protein engineering to improve both editing efficiency and targeting accuracy of Acidaminococcus sp. Cas12a (AsCas12a) and Lachnospiraceae bacterium Cas12a (LbCas12a).
resultsWe created new AsCas12a variant termed "AsCas12a-Plus" with increased activity (1.5~2.0-fold improvement) and specificity (reducing off-targets from 29 to 23 and specificity index increased from 92% to 94% with 33 sgRNAs), and this property was retained in multiplex editing and transcriptional activation. When used to disrupt the oncogenic BRAF
conclusionsOur discoveries provide alternative AsCas12a and LbCas12a variants with high specificity and activity, which expand the gene-editing toolbox and can be more suitable for clinical applications.
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