Fig. 3. Reprogrammed tracrRNAs co-opt RNA transcripts to guide different Cas9 orthologs.
(A) Design of reprogrammed tracrRNAs (Rptrs) utilized by CjeCas9. W = A or T. PAM, yellow circle. (B) Toleration of mutations to the RNA duplex of an sgRNA that preserve secondary structure in TXTL. (C) Enhancing DNA targeting by less functional or non-functional ncrRNAs by converting the tracrRNA into a Rptr in TXTL. (D) Sequence-specific DNA targeting in TXTL using Rptrs compatible with three different Cas9 nucleases. Values in (B) to (D) represent the mean and standard deviation from four replicates in TXTL. *p < 0.01; **p < 0.001. n.s., not significant.