Details

Autor(en) / Beteiligte

• Rauch, Benjamin J.
• Silvis, Melanie R.
• Hultquist, Judd F.
• Waters, Christopher S.
• McGregor, Michael J.
• Krogan, Nevan J.
• Bondy-Denomy, Joseph

Titel

Inhibition of CRISPR-Cas9 with Bacteriophage Proteins

Ist Teil von

• Cell, 2017-01, Vol.168 (1-2), p.150-158.e10

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2017

Quelle

MEDLINE

Beschreibungen/Notizen

• Bacterial CRISPR-Cas systems utilize sequence-specific RNA-guided nucleases to defend against bacteriophage infection. As a countermeasure, numerous phages are known that produce proteins to block the function of class 1 CRISPR-Cas systems. However, currently no proteins are known to inhibit the widely used class 2 CRISPR-Cas9 system. To find these inhibitors, we searched cas9-containing bacterial genomes for the co-existence of a CRISPR spacer and its target, a potential indicator for CRISPR inhibition. This analysis led to the discovery of four unique type II-A CRISPR-Cas9 inhibitor proteins encoded by Listeria monocytogenes prophages. More than half of L. monocytogenes strains with cas9 contain at least one prophage-encoded inhibitor, suggesting widespread CRISPR-Cas9 inactivation. Two of these inhibitors also blocked the widely used Streptococcus pyogenes Cas9 when assayed in Escherichia coli and human cells. These natural Cas9-specific “anti-CRISPRs” present tools that can be used to regulate the genome engineering activities of CRISPR-Cas9. [Display omitted] •Bacteriophage anti-CRISPR proteins inactivate Listeria monocytogenes CRISPR-Cas9•Half of L. monocytogenes isolates possess inhibited CRISPR-Cas9 systems•AcrIIA2 and AcrIIA4 prevent target binding by dCas9 in bacteria•AcrIIA2 and AcrIIA4 inhibit Cas9-mediated gene editing in human cells Four CRISPR-Cas9 inhibitor proteins encoded by Listeria monocytogenes prophages prevent Cas9 binding and gene editing in bacteria and human cells, including currently the most widely used Cas9 from Streptococcus pyogenes.