A Cas9 with PAM recognition for adenine dinucleotides
Name
s41467-020-16117-8.pdf
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1.45 MB
Format
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Checksum (MD5)
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Author(s)
Chatterjee, Pranam
Lee, Jooyoung
Nip, Lisa
Koseki, Sabrina R. T.
Tysinger, Emma
Sontheimer, Erik J.
Jacobson, Joseph
Jakimo, Noah
Date Issued
May 2020
Journal
Nature Communications
Publisher
Springer Science and Business Media LLC
Citation
Chatterjee, Pranam et al. "A Cas9 with PAM recognition for adenine dinucleotides." Nature Communications 11, 1 (May 2020): 2474 © 2020 Springer Nature
Version
Final published version
Abstract
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′-NAAN-3′ 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.
MIT Department
Massachusetts Institute of Technology. Center for Bits and Atoms
Massachusetts Institute of Technology. Media Laboratory
Terms of Use
Creative Commons Attribution 4.0 International license
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DOI of Published Version
https://doi.org/10.1038/s41467-020-16117-8
