Figure 3.

AcrIIA24–32 inhibit the Streptococcus type II-A Cas9 orthologs in vitro. (A) Schematic overview of DNA cleavage assays, in which Cas9 RNP was used to target a DNA substrate in the presence of Acr proteins. DNA cleavage assays using SpyCas9 (B), St3Cas9 (C) and St1Cas9 (D) RNPs to target a linearized plasmid DNA in the presence or absence of Acr proteins. Acr subtypes and numbers are indicated. A, AcrIIA; C, AcrIIC. Hollow arrowheads indicate the uncleaved linearized plasmid DNA. Solid arrowheads indicate cleaved products. The gel images shown for DNA cleavage assays are representative of three independent replicates.