Fig. 1.

Cas9:tracrRNA complexed with RNA or hybrid crXNAs are nuclease competent. a Design of the crRNA, crDNA and crHyb sequence for GFP target site 1 (GFP cr1). DNA shown in uppercase black, RNA shown in lowercase red. b Cas9, tracrRNA and crXNA molecules were incubated with linear double-stranded DNA harbouring the GFP target sequence. Reaction products were analysed by agarose gel electrophoresis. The expected undigested and digested fragments are exemplified on the right. c Semiquantitative analysis of the cleavage of the linear DNA fragment by crDNA (blue)-, crHyb (black)- and crRNA (red)-guided Cas9:tracrRNA complexes with three independent guide sequences. Bars show mean + SD, n = 3–7