Fig. 6
From: Structural basis for the promiscuous PAM recognition by Corynebacterium diphtheriae Cas9
Protospacer adjacent motif (PAM) recognition by Corynebacterium diphtheriae Cas9 (CdCas9). a Binding of the PAM duplex to CdCas9. b Schematics of the PAM recognition by CdCas9. Hydrogen bonds and hydrophobic interactions are depicted by black and red dashed lines, respectively. c Recognition of the GGGTAAT PAM (stereo view). Hydrogen bonds are shown as dashed lines. d Effects of the mutations on the PAM-interacting residues. The linearized plasmid target with the GGGAAAC PAM was incubated with the wild-type or the mutants of CdCas9, together with the single-guide RNA (sgRNA22), at 37 °C for 0.5 and 5 min. FPL, F1011A/P1043A/L1046A; FPLK, F1011A/K1015A/P1043A/L1046A. e Functional importance of the second PAM nucleotide. The linearized plasmid targets bearing the different PAMs were incubated with CdCas9-sgRNA22 at 37 °C for 0.5 and 5 min. Error bars represent s.d. from n = 3 replicates
