RNA Therapeutics Institute
Bioinformatics | Computational Biology | Enzymes and Coenzymes | Genetic Phenomena | Genetics and Genomics | Nucleic Acids, Nucleotides, and Nucleosides
CRISPR-associated (Cas) DNA-endonucleases are remarkably effective tools for genome engineering, but have limited target ranges due to their protospacer adjacent motif (PAM) requirements. We demonstrate a critical expansion of the targetable sequence space for a type II-A CRISPR-associated enzyme through identification of the natural 5[Formula: see text]-NAAN-3[Formula: see text] PAM preference of Streptococcus macacae Cas9 (SmacCas9). To achieve efficient editing activity, we graft the PAM-interacting domain of SmacCas9 to its well-established ortholog from Streptococcus pyogenes (SpyCas9), and further engineer an increased efficiency variant (iSpyMac) for robust genome editing activity. We establish that our hybrids can target all adenine dinucleotide PAM sequences and possess robust and accurate editing capabilities in human cells.
CRISPR-Cas9 genome editing, Computational biology and bioinformatics, Synthetic biology
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DOI of Published Version
Chatterjee P, Lee J, Nip L, Koseki SRT, Tysinger E, Sontheimer EJ, Jacobson JM, Jakimo N. A Cas9 with PAM recognition for adenine dinucleotides. Nat Commun. 2020 May 18;11(1):2474. doi: 10.1038/s41467-020-16117-8. PMID: 32424114; PMCID: PMC7235249. Link to article on publisher's site
Chatterjee P, Lee J, Nip L, Koseki SR, Tysinger E, Sontheimer EJ, Jacobson JM, Jakimo N. (2020). A Cas9 with PAM recognition for adenine dinucleotides. Open Access Articles. https://doi.org/10.1038/s41467-020-16117-8. Retrieved from https://escholarship.umassmed.edu/oapubs/4255
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.