Fig. 6.

The StCas9 system functions on targets integrated in 293T genome. a Detection of SSA-mediated repair frequencies for integrated reporter constructs. 293T.SP1.PAM5* and 293T.SP1.P5M4* represent cells transfected with corresponding sgRNA and hStCas9 expression vectors. b Mutations detected in the integrated reporter constructs caused by NHEJ repair pathway. 2 of 48 and 3 of 44 for the targets SP1.PAM5 and SP1.P5M4, respectively. c Comparison of wild type (SP1.gRNA.WT) and optimized (SP1.gRNA.Opti) designs for guiding the StCas9 activity on integrated reporter targets in 293T cells