Temperature-Responsive Competitive Inhibition of CRISPR-Cas9

Jiang, Fuguo; Liu, Jun-Jie; Osuna, Beatriz A.; Xu, Michael; Berry, Joel D.; Rauch, Benjamin J.; Nogales, Eva; Bondy-Denomy, Joseph; Doudna, Jennifer A.

doi:10.1016/j.molcel.2018.11.016

Temperature-Responsive Competitive Inhibition of CRISPR-Cas9

Journal Article · Wed Dec 26 19:00:00 EST 2018 · Molecular Cell

DOI:https://doi.org/10.1016/j.molcel.2018.11.016· OSTI ID:1604673

Jiang, Fuguo ^[1]; Liu, Jun-Jie ^[1]; Osuna, Beatriz A. ^[2]; Xu, Michael ^[1]; Berry, Joel D. ^[2]; Rauch, Benjamin J. ^[2]; Nogales, Eva ^[3]; Bondy-Denomy, Joseph ^[2]; Doudna, Jennifer A. ^[4]

Univ. of California, Berkeley, CA (United States)
Univ. of California, San Francisco, CA (United States)
Univ. of California, Berkeley, CA (United States); Howard Hughes Medical Inst., Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Univ. of California, Berkeley, CA (United States); Innovative Genomics Inst., Berkeley, CA (United States); Howard Hughes Medical Inst., Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Gladstone Inst., San Francisco, CA (United States)

CRISPR-Cas immune systems utilize RNA-guided nucleases to protect bacteria from bacteriophage infection. Bacteriophages have in turn evolved inhibitory "anti-CRISPR" (Acr) proteins, including six inhibitors (AcrIIA1-AcrIIA6) that can block DNA cutting and genome editing by type II-A CRISPR-Cas9 enzymes. We show here that AcrIIA2 and its more potent homolog, AcrIIA2b, prevent Cas9 binding to DNA by occluding protein residues required for DNA binding. Cryo-EM-determined structures of AcrIIA2 or AcrIIA2b bound to S. pyogenes Cas9 reveal a mode of competitive inhibition of DNA binding that is distinct from other known Acrs. Differences in the temperature dependence of Cas9 inhibition by AcrIIA2 and AcrIIA2b arise from differences in both inhibitor structure and the local inhibitor-binding environment on Cas9. Finally, these findings expand the natural toolbox for regulating CRISPR-Cas9 genome editing temporally, spatially, and conditionally.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC); Defense Advanced Research Projects Agency (DARPA)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1604673

Journal Information:: Molecular Cell, Journal Name: Molecular Cell Journal Issue: 3 Vol. 73; ISSN 1097-2765

Publisher:: Cell Press - ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
AcrllA2
CRISPR-Cas
CRISPR-Cas9
Cas9
Cas9 inhibitors
PAM blockage
anti-CRISPR
bacteriophage
genome editing
temperature-dependent inhibition

Temperature-Responsive Competitive Inhibition of CRISPR-Cas9

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References (55)

Cited By (13)

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