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. 2020 Apr 14;48(9):4698–4708. doi: 10.1093/nar/gkaa219

Figure 2.

Figure 2.

Inhibition of SpyCas9 and SauCas9 by newly discovered Acr candidates. (A) In vitro cleavage of dsDNA by SpyCas9 in the absence or presence of a 50-fold excess of AcrIIA4 (positive control) and each Acr candidate. (B) In vitro cleavage of dsDNA by SpyCas9 in the presence of increasing concentrations of (left to right) BSA (negative control), AcrIIA4 (positive control), ML1 and ML8 (Acr:RNP 0.1-, 1-, 2-,10-, 50- and 100-fold excess from left to right). (C) In vitro cleavage of dsDNA by SauCas9 in the absence or presence of a 25-fold excess of each Acr candidate. (D) In vitro cleavage of dsDNA by SauCas9 in the presence of increasing concentrations of (left to right) BSA (negative control), AcrllA5 (positive control, Acr:RNP 0.1-, 1-, 2-, 4-, 8- and 10-fold excess from left to right), ML3 and ML8 (Acr:RNP 0.1-, 1-, 2-,10-, 50- and 100-fold excess from left to right). Uncropped gel images for panels B and D are shown in Supplementary Figures S3 and S4.