Fig. 2.

Cas9:tracrRNA can function with a crXNA composed of 88% DNA and crHyb show enhanced specificity. a RNA nucleosides within the crHyb for GFP target 1 were sequentially converted to DNA and activity assessed by DNA fragmentation assay. Nucleotides coloured as in Fig. 1a. b A single base C > A mismatch was introduced into crRNA and crHyb molecules and activity assessed. All bars show mean + SD, n = 3–13. c Kinetics of template degradation driven by Cas9:tracrRNA complexed with crHyb (black) or crRNA (red), either with perfect complementary (left) or with a single base-pair mismatch (right). Graphs show normalised template abundance (%) over time, with Cas9 protein at 5 nM (solid line), 2.5 nM (long dash line), 1.25 nM (dot dash line) or 0.625 nM (dotted line). Error bars show SD of 3–4 independent measurements