Fig. 1

In vitro cleavage activities of Corynebacterium diphtheriae Cas9 (CdCas9). a In vitro cleavage activities of CdCas9 with the 20–24-nt guide single-guide RNAs (sgRNAs). The linearized plasmid target bearing the GGGAAAC protospacer adjacent motif (PAM) was incubated with the CdCas9-sgRNA complex at 37 °C for 0.5, 1, 2, and 5 min, and the cleavage products were then analyzed by a MultiNA microchip electrophoresis system. b In vitro cleavage activities of CdCas9 and Streptococcus pyogenes Cas9 (SpCas9). CdCas9 and SpCas9 were programmed with their 22- and 20-nt guide sgRNAs, respectively. The linearized plasmid target bearing the GGGAAAC PAM was incubated with the Cas9-sgRNA complex at 37 °C for 0.25, 0.5, 1, 2, and 5 min. c Motif obtained from the in vitro PAM identification assay. d PAM preference of CdCas9. 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