ArticleMolecular therapy : the journal of the American Society of Gene Therapy2022
Cas12a variants designed for lower genome-wide off-target effect through stringent PAM recognition.
Article in Molecular therapy : the journal of the American Society of Gene Therapy, 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, 37 citations in OpenAlex.
- Mismatch-Driven CRISPR/Cas12a Biosensing of UV-Induced DNA Lesions for Environmental Solar Exposure Surveillance.Environmental science & technology · 2026Article
- High-fidelity molecular decoding through tile-guided in situ self-assembly.Science advances · 2026Article
- Mechanistic basis for improved activity of Engineered AsCas12a.Communications biology · 2026Article
- CRISPR/Cas9 in cancer therapy: clinical translation, mechanistic strategies, and therapeutic directions.Frontiers in oncology · 2026Review
- Understanding the stability landscape of LbCas12a by deep analysis of stabilizing mutations and mutation combinations.Protein science : a publication of the Protein Society · 2025Article
- A CRISPR Cas12a/Cpf1 strategy to facilitate robust multiplex gene editing in Aspergillus Niger.Fungal biology and biotechnology · 2025Article
- Emerging tools in plant genome editing.Frontiers in genome editing · 2025Review
- Off-target sequence variations driven by the intrinsic properties of the Cas-sgRNA-DNA complex in genome editing.PloS one · 2025Article
- State of the art CRISPR-based strategies for cancer diagnostics and treatment.Biomarker research · 2024Review
- Recent Advances in CRISPR/Cas System-Based Biosensors for the Detection of Foodborne Pathogenic Microorganisms.Micromachines · 2024Review
- Review
- CRISPR-enabled point-of-care genotyping for APOL1 genetic risk assessment.EMBO molecular medicine · 2024Article
- High-fidelity CRISPR/Cas12a dual-crRNA screening reveals novel synergistic interactions in hepatocellular carcinoma.Clinical and translational medicine · 2024Article
- CoHIT: a one-pot ultrasensitive ERA-CRISPR system for detecting multiple same-site indels.Nature communications · 2024Article
- Multiplexed in-situ mutagenesis driven by a dCas12a-based dual-function base editor.Nucleic acids research · 2024Article
- A high-fidelity DNAzyme-assisted CRISPR/Cas13a system with single-nucleotide resolved specificity.Chemical science · 2024Article
- Engineering of a compact, high-fidelity EbCas12a variant that can be packaged with its crRNA into an all-in-one AAV vector delivery system.PLoS biology · 2024Article
- Engineering of Cas12a nuclease variants with enhanced genome-editing specificity.PLoS biology · 2024Article
- Discovery of Diverse CRISPR-Cas Systems and Expansion of the Genome Engineering Toolbox.Biochemistry · 2023Review
- Cas9-mediated knockout of Ndrg2 enhances the regenerative potential of dendritic cells for wound healing.Nature communications · 2023Article
Corrections and comments
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Authors and funding
11 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Cas12a is an RNA-guided endonuclease that has been widely used for convenient multiplex gene editing with low off-target effects. To minimize off-targeting in gene editing, we engineered a variant of LbCas12a (termed Lb-K538R) with more stringent PAM recognition, lower off-targeting capability, and similar editing efficiency in vivo compared with LbCas12a. We also demonstrated that Lb2Cas12a from Lachnospiraceae bacterium MA2020 has extensive gene-editing activities in mammalian cells. Similar to Lb-K538R, the designed Lb2Cas12a variant (termed Lb2-K518R) not only had a more stringent PAM sequence change from YYN to TYN (Y is T or C, N is A, T, C, or G), but also displayed lower off-target effects, thereby enabling more potential target site selections with low off-targeting than the common TTTV (V is A, G, or C) PAM. To determine whether this type of mutation at the homologous position had similar effects in other Cas12a, As-K548R was evaluated. Based on the results of the genome-wide off-target test, As-K548R displayed lower off-target effects. Collectively, our findings indicate that the Cas proteins could be designed to be stringent in PAM recognition to reduce their off-target effects, which suggests a promising and practical approach for minimizing off-targets effects in genome editing.
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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.