ArticlePLoS biology2022
Identification of SaCas9 orthologs containing a conserved serine residue that determines simple NNGG PAM recognition.
Article in PLoS biology, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- GenomePAM directs PAM characterization and engineering of CRISPR-Cas nucleases using mammalian genome repeats.Nature biomedical engineering · 2026Article
- Optimized in vivo base editing restores auditory function in a DFNA15 mouse model.Nature communications · 2025Article
- Small RNA Toxin-Assisted Evolution of GC-Preferred ErCas12a for Enhanced Genome Targeting Range.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- GenomePAM directs PAM characterization and engineering of CRISPR-Cas nucleases using mammalian genome repeats.Research square · 2025Article
- Engineering of SauriCas9 with enhanced specificity.Molecular therapy. Nucleic acids · 2025Article
- Correction: Identification of SaCas9 orthologs containing a conserved serine residue that determines simple NNGG PAM recognition.PLoS biology · 2025Article
- Characterization of NiCas12b for In Vivo Genome Editing.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
- Efficient and markerless gene integration with SlugCas9-HF in Kluyveromyces marxianus.Communications biology · 2024Article
- Engineering of a high-fidelity Cas12a nuclease variant capable of allele-specific editing.PLoS biology · 2024Article
- Phage-assisted evolution of compact Cas9 variants targeting a simple NNG PAM.Nature chemical biology · 2024Article
- CRISPR/Cas9 Landscape: Current State and Future Perspectives.International journal of molecular sciences · 2023Review
- Engineering aACS synthetic biology · 2023Article
- Full-Length Model of SaCas9-sgRNA-DNA Complex in Cleavage State.International journal of molecular sciences · 2023Article
Corrections and comments
- Erratum issued
Authors and funding
11 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Due to different nucleotide preferences at target sites, no single Cas9 is capable of editing all sequences. Thus, this highlights the need to establish a Cas9 repertoire covering all sequences for efficient genome editing. Cas9s with simple protospacer adjacent motif (PAM) requirements are particularly attractive to allow for a wide range of genome editing, but identification of such Cas9s from thousands of Cas9s in the public database is a challenge. We previously identified PAMs for 16 SaCas9 orthologs. Here, we compared the PAM-interacting (PI) domains in these orthologs and found that the serine residue corresponding to SaCas9 N986 was associated with the simple NNGG PAM requirement. Based on this discovery, we identified five additional SaCas9 orthologs that recognize the NNGG PAM. We further identified three amino acids that determined the NNGG PAM requirement of SaCas9. Finally, we engineered Sha2Cas9 and SpeCas9 to generate high-fidelity versions of Cas9s. Importantly, these natural and engineered Cas9s displayed high activities and distinct nucleotide preferences. Our study offers a new perspective to identify SaCas9 orthologs with NNGG PAM requirements, expanding the Cas9 repertoire.
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