Chapter 5 —— 129 —— Supplementary Figures Supplementary Figure S1. Testing selector AAV-based genome editing at AAVS1. (A) Diagram of the experimental setup. HeLa cells transfected with plasmids expressing Cas9 and mCherry or inactive dCas9 and mCherry are transduced with the selector AAVHRS1.A1 donor and, after sub-culturing in the presence or absence of ouabain, are subjected to EGFP- and mCherry-directed flow cytometry. HeLa cells only exposed to AAV-HRS1.A1 served as a control. (B) Quantification of transfection efficiency and selector AAV donor delivery. The transfection efficiency and AAV-HRS1.A1 donor delivery were determined by mCherry- and EGFP-directed flow cytometry at 3 days post-transduction (left and central graphs, respectively). The frequencies of transfected and transduced cells are also plotted (right graph). (C) Quantification of selector AAV-based DNA editing. The AAV stable transduction levels, serving as a proxy for genome editing frequencies, were determined at 20 days post-transduction by EGFP-directed flow cytometry. The cross indicates complete cell death in ouabain-treated cultures exposed exclusively to AAVHRS1.A1.
RkJQdWJsaXNoZXIy MTk4NDMw