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A Cas9 with PAM recognition for adenine dinucleotides

Publication ,  Journal Article
Chatterjee, P; Lee, J; Nip, L; Koseki, SRT; Tysinger, E; Sontheimer, EJ; Jacobson, JM; Jakimo, N
Published in: Nature Communications
May 18, 2020
Published version (DOI) Publisher Link

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 Cas9 (SmacCas9). To achieve efficient editing activity, we graft the PAM-interacting domain of SmacCas9 to its well-established ortholog from (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.

Duke Scholars

Pranam D. Chatterjee
Author Pranam D. Chatterjee Biomedical Engineering
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Published In

Nature Communications

DOI

10.1038/s41467-020-16117-8

EISSN

2041-1723

Publication Date

May 18, 2020

Volume

11

Issue

1

Publisher

Springer Science and Business Media LLC
 

Citation

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Chatterjee, P., Lee, J., Nip, L., Koseki, S. R. T., Tysinger, E., Sontheimer, E. J., … Jakimo, N. (2020). A Cas9 with PAM recognition for adenine dinucleotides. Nature Communications, 11(1). https://doi.org/10.1038/s41467-020-16117-8
Chatterjee, Pranam, Jooyoung Lee, Lisa Nip, Sabrina R. T. Koseki, Emma Tysinger, Erik J. Sontheimer, Joseph M. Jacobson, and Noah Jakimo. “A Cas9 with PAM recognition for adenine dinucleotides.” Nature Communications 11, no. 1 (May 18, 2020). https://doi.org/10.1038/s41467-020-16117-8.
Chatterjee P, Lee J, Nip L, Koseki SRT, Tysinger E, Sontheimer EJ, et al. A Cas9 with PAM recognition for adenine dinucleotides. Nature Communications. 2020 May 18;11(1).
Chatterjee, Pranam, et al. “A Cas9 with PAM recognition for adenine dinucleotides.” Nature Communications, vol. 11, no. 1, Springer Science and Business Media LLC, May 2020. Crossref, doi:10.1038/s41467-020-16117-8.
Chatterjee P, Lee J, Nip L, Koseki SRT, Tysinger E, Sontheimer EJ, Jacobson JM, Jakimo N. A Cas9 with PAM recognition for adenine dinucleotides. Nature Communications. Springer Science and Business Media LLC; 2020 May 18;11(1).

Published In

Nature Communications

DOI

10.1038/s41467-020-16117-8

EISSN

2041-1723

Publication Date

May 18, 2020

Volume

11

Issue

1

Publisher

Springer Science and Business Media LLC