ArticleNucleic acids research2020
Improved LbCas12a variants with altered PAM specificities further broaden the genome targeting range of Cas12a nucleases.
Article in Nucleic acids research, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 78 papers.
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Who cites it
78 citing papers in PubMed.
- Establishment of an efficient and PAM-relaxed LbCas12a genome editing tool in plants.The New phytologist · 2026Article
- Optimized tRNA processing and TREX2-SpCas9 fusion enable high-efficiency multiplex genome editing in plants.Plant communications · 2026Article
- CRISPR diagnostics: from trans-nuclease activity to cancer diagnosis.Cell & bioscience · 2026Review
- A rapid and highly sensitive CRISPR-Cas12a ortholog-assisted assay for genotyping of myostatin knockout pigs.Cell & bioscience · 2026Article
- Structure-Guided Engineering of a Cas12i Nuclease Unlocks Near-PAMless Genome Editing.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- From Lab to Field: CRISPRing Major CultivatedInternational journal of molecular sciences · 2026Review
- Benchmarking the PAM compatibility of Cas12a variants for high-throughput yeast genetic variant engineering.Applied and environmental microbiology · 2025Article
- Development and validation of PmMAD7 for efficient gene editing in Penaeus monodon.BMC biotechnology · 2025Article
- Application of engineered CRISPR/Cas12a variants with altered protospacer adjacent motif specificities for the detection of isoniazid resistance mutations inMicrobiology spectrum · 2025Article
- Understanding the stability landscape of LbCas12a by deep analysis of stabilizing mutations and mutation combinations.Protein science : a publication of the Protein Society · 2025Article
- Small RNA Toxin-Assisted Evolution of GC-Preferred ErCas12a for Enhanced Genome Targeting Range.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Directed evolution expands CRISPR-Cas12a genome-editing capacity.Nucleic acids research · 2025Article
- Never-germinating Arabidopsis seeds with LbCas12a-induced mutations in 6 clade A type 2C protein phosphatase genes.Plant physiology · 2025Article
- Dual Cas12a and multiplex crRNA CRISPR strategy ultrasensitive detection novel circRNA biomarker for the diagnosis of ovarian cancer.BMC cancer · 2025Article
- Post-cleavage target residence determines asymmetry in non-homologous end joining of Cas12a-induced DNA double strand breaks.Genome biology · 2025Article
- Highly parallel profiling of the activities and specificities of Cas12a variants in human cells.Nature communications · 2025Article
- PAM-Independent CRISPR-Cas12a System for Specific Assays of Single Nucleotide Variants.JACS Au · 2025Article
- Insight into crRNA Processing inInternational journal of molecular sciences · 2025Article
- PathoGD: an integrative genomics approach to primer and guide RNA design for CRISPR-based diagnostics.Communications biology · 2025Article
- PAM-relaxed and temperature-tolerant CRISPR-Mb3Cas12a single transcript unit systems for efficient singular and multiplexed genome editing in rice, maize, and tomato.Plant biotechnology journal · 2025Article
18 more citing papers are in PubMed but not listed here.
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Authors and funding
11 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The widespread use of Cas12a (formerly Cpf1) nucleases for genome engineering is limited by their requirement for a rather long TTTV protospacer adjacent motif (PAM) sequence. Here we have aimed to loosen these PAM constraints and have generated new PAM mutant variants of the four Cas12a orthologs that are active in mammalian and plant cells, by combining the mutations of their corresponding RR and RVR variants with altered PAM specificities. LbCas12a-RVRR showing the highest activity was selected for an in-depth characterization of its PAM preferences in mammalian cells, using a plasmid-based assay. The consensus PAM sequence of LbCas12a-RVRR resembles a TNTN motif, but also includes TACV, TTCV CTCV and CCCV. The D156R mutation in improved LbCas12a (impLbCas12a) was found to further increase the activity of that variant in a PAM-dependent manner. Due to the overlapping but still different PAM preferences of impLbCas12a and the recently reported enAsCas12a variant, they complement each other to provide increased efficiency for genome editing and transcriptome modulating applications.
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