U2OS cells were transfected with equal amounts of Edit-R hCMV-mKate2-Cas9 Expression plasmid (Cat #U-004100-120) and Edit-R Human PPIB Lentiviral sgRNA plasmid in a 96-well format. Transient transfections were done with increasing amounts of total DNA (50 to 200 ng) and DharmaFECT kb transfection reagent (Cat #T-2006-01), 0.2 to 0.8 µL per well. The percentage of gene editing was estimated 72 hours after transfection by DNA mismatch detection assay using T7EI with gel densitometry.
HEK293T cells were transfected with 1 µg SMARTvector Human AKT1 Lentiviral shRNA plasmid (Cat #V3SH11240-224972267) in a 24-well format. Transient transfections were done with DharmaFECT kb transfection reagent (Cat #T-2006-01) at 3 µL per well. Images were taken 48 hours after transfection.