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. 2015 Oct 9;4(11):1217–1225. doi: 10.1021/acssynbio.5b00132

Figure 1.

Figure 1

CRISPR-directed SSBs are not lethal to K12 E. coli. (a) A schematic representation of sgRNA guided Cas9s targeting a chromosomally integrated GFP reporter. The sgRNA guide sequence is visualized adjacent to its target sequence. The protospacer adjacent motif (PAM) sequence is highlighted in red. Arrows denote where DNA cleavage occurs by two Cas9 mutants, respectively. Cas9D10A cleaves the target DNA strand of the sgRNA and Cas9H840A cleaves the nontarget strand. (b) Transformation efficiencies of sg(C1) (gray) or untargeted sg(−) (white) in K12 E. coli containing Cas9WT, CasD10A, Cas9H840A or dCas9. Error bars represent ± standard deviation (n = 3).