Fig. 1: In vitro cleavage activity.
From: Structure and engineering of Brevibacillus laterosporus Cas9
a In vitro DNA cleavage activities of BlCas9 with the 20–23 nt guide sgRNAs toward three different target sequences (Targets 1–3). The linearized plasmid target bearing the T3CCCAA (Target 1) or T3CCGAA (Targets 2 and 3) PAM was incubated with the BlCas9–sgRNA complex at 37 °C for 0.5, 1, 2, and 5 min. The cleavage products were then analyzed by a MultiNA microchip electrophoresis system. Data are mean ± s.d. (n = 3). b, c Sequence logo (b) and 2D profile (c) of the BlCas9 PAM obtained from the PAM identification assay. d In vitro DNA cleavage activities of BlCas9 with the 22-nt guide sgRNA toward DNA targets (Target 1) with different PAMs. The linearized plasmid targets were incubated with the BlCas9–sgRNA complex at 37 °C for 2 and 5 min. Data are mean ± s.d. (n = 3).
