ArticleNucleic acids research2022
Mechanics of CRISPR-Cas12a and engineered variants on λ-DNA.
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 13 papers.
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Who cites it
13 citing papers in PubMed, 24 citations in OpenAlex.
- Size and sequence variation of the Lid of the Cas12a nuclease domain results in a nickase phenotype.Nucleic acids research · 2026Article
- Advances in biotechnological methods for genetic and metabolic engineering in Methylomonas sp. DH-1.Journal of biological engineering · 2026Review
- Dynamic basis of supercoiling-dependent DNA interrogation by Cas12a via R-loop intermediates.Nature communications · 2025Article
- PAM-adjacent DNA flexibility tunes CRISPR-Cas12a off-target binding.Scientific reports · 2025Article
- Cas12a domain flexibility guides R-loop formation and forces RuvC resetting.Molecular cell · 2024Article
- Live-cell imaging reveals the trade-off between target search flexibility and efficiency for Cas9 and Cas12a.Nucleic acids research · 2024Article
- An alpha-helical lid guides the target DNA toward catalysis in CRISPR-Cas12a.Nature communications · 2024Article
- Utilization of nicking properties of CRISPR-Cas12a effector for genome editing.Scientific reports · 2024Article
- Transposons and CRISPR: Rewiring Gene Editing.Biochemistry · 2023Article
- Force-Triggered Self-Destructive Hydrogels.Advanced materials (Deerfield Beach, Fla.) · 2023Article
- Targeting miRNA by CRISPR/Cas in cancer: advantages and challenges.Military Medical Research · 2023Review
- Systematic optimization of Cas12a base editors in wheat and maize using the ITER platform.Genome biology · 2023Article
- Targeted Modification of Mammalian DNA by a Novel Type V Cas12a Endonuclease fromInternational journal of molecular sciences · 2022Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Cas12a is an RNA-guided endonuclease that is emerging as a powerful genome-editing tool. Here, we selected a target site on bacteriophage λ-DNA and used optical tweezers combined with fluorescence to provide mechanistic insight into wild type Cas12a and three engineered variants, where the specific dsDNA and the unspecific ssDNA cleavage are dissociated (M1 and M2) and a third one which nicks the target DNA (M3). At low forces wtCas12a and the variants display two main off-target binding sites, while on stretched dsDNA at higher forces numerous binding events appear driven by the mechanical distortion of the DNA and partial matches to the crRNA. The multiple binding events onto dsDNA at high tension do not lead to cleavage, which is observed on the target site at low forces when the DNA is flexible. In addition, activity assays also show that the preferential off-target sites for this crRNA are not cleaved by wtCas12a, indicating that λ-DNA is only severed at the target site. Our single molecule data indicate that the Cas12a scaffold presents singular mechanical properties, which could be used to generate new endonucleases with biomedical and biotechnological applications.
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