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. 2017 Jun 29;45(14):8411–8422. doi: 10.1093/nar/gkx572

Figure 5.

Figure 5.

Attempted Cas9-PB mediated gene targeting of HPRT. (A) Surveyor assay using the five gRNAs as depicted in Figure 4A with catalytically active hSpCas9 fused to PB and co-transfected with the PB-SB-SA-βGeo plasmid. (B) G418 resistant colonies produced after co-transfection with the PB-SB-SA-βGeo plasmid with Cas9-PB and the varying gRNAs. ANOVA followed by Bonferroni post-test analysis revealed no difference between conditions (N = 4, ±SEM). (C) G418 resistant colonies produced after co-transfection with the PB-SB-SA-βGeo plasmid with Cas9 and PB on separate plasmids with or without the varying gRNAs. ANOVA followed by Bonferroni post-test analysis revealed no difference between conditions (N = 4, ±SEM). (D) Cas9-PB colony number after replating G418 resistant cells into and selecting with 6-TG. ANOVA with Bonferroni post-test revealed E2 and E3 to be significantly different from no gRNA (N = 4, ±SEM). (E) Cas9 + PB (supplied separately) colony number after replating G418 resistant cells into and selecting with 6-TG. ANOVA with Bonferroni post-test revealed E2 and E3 to be significantly different from no gRNA (N = 4, ±SEM).