ArticleMolecular cell2016
Structural Plasticity of PAM Recognition by Engineered Variants of the RNA-Guided Endonuclease Cas9.
Article in Molecular cell, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 103 papers, 1 of them a synthesis that pooled it.
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Who cites it
103 citing papers in PubMed, 1 synthesis or guideline pooled it.
- An overview and metanalysis of machine and deep learning-based CRISPR gRNA design tools.RNA biology · 2020Pooled it
- Treatment of a severe vascular disease using a bespoke CRISPR-Cas9 base editor in mice.Nature biomedical engineering · 2026Article
- Successful In Vitro Modification of theCells · 2026Article
- Review
- Engineered Un1Cas12f1 for multiplex genome editing with enhanced activity and targeting scope.Nature communications · 2026Article
- Customizing CRISPR-Cas PAM specificity with protein language models.Nature biotechnology · 2026Article
- CRISPR/Cas-mediated genome editing: playing a versatile role in mitigating the challenges of sustainable rice improvement.3 Biotech · 2025Review
- Design Rules for Expanding PAM Compatibility in CRISPR-Cas9 from the VQR, VRER and EQR variants.bioRxiv : the preprint server for biology · 2025Article
- CRISPR-based functional genomics tools in vertebrate models.Experimental & molecular medicine · 2025Review
- Article
- CRISPR/Cas system-guided plasmid mutagenesis without sequence restriction.Fundamental research · 2025Article
- TOP-SECRETS enables Cas9 nucleases to discriminate SNVs outside of PAMs.bioRxiv : the preprint server for biology · 2025Article
- Predicting adenine base editing efficiencies in different cellular contexts by deep learning.Genome biology · 2025Article
- DNA targeting by compact Cas9d and its resurrected ancestor.Nature communications · 2025Article
- Engineering of CRISPR-Cas PAM recognition using deep learning of vast evolutionary data.bioRxiv : the preprint server for biology · 2025Article
- Gene Editing: An Effective Tool for the Future Treatment of Kidney Disease.Journal of inflammation research · 2025Review
- Harnessing the evolving CRISPR/Cas9 for precision oncology.Journal of translational medicine · 2024Review
- AlPaCas: allele-specific CRISPR gene editing through a protospacer-adjacent-motif (PAM) approach.Nucleic acids research · 2024Article
- PAM-flexible Engineered FnCas9 variants for robust and ultra-precise genome editing and diagnostics.Nature communications · 2024Article
- Discovery and structural mechanism of DNA endonucleases guided by RAGATH-18-derived RNAs.Cell research · 2024Article
43 more citing papers are in PubMed but not listed here.
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3 authors.
Funding
Abstract
The RNA-guided endonuclease Cas9 from Streptococcus pyogenes (SpCas9) forms the core of a powerful genome editing technology. DNA cleavage by SpCas9 is dependent on the presence of a 5'-NGG-3' protospacer adjacent motif (PAM) in the target DNA, restricting the choice of targetable sequences. To address this limitation, artificial SpCas9 variants with altered PAM specificities have recently been developed. Here we report crystal structures of the VQR, EQR, and VRER SpCas9 variants bound to target DNAs containing their preferred PAM sequences. The structures reveal that the non-canonical PAMs are recognized by an induced fit mechanism. Besides mediating sequence-specific base recognition, the amino acid substitutions introduced in the SpCas9 variants facilitate conformational remodeling of the PAM region of the bound DNA. Guided by the structural data, we engineered a SpCas9 variant that specifically recognizes NAAG PAMs. Taken together, these studies inform further development of Cas9-based genome editing tools.
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