Fig. 3
From: Enhancement of CRISPR-Cas9 induced precise gene editing by targeting histone H2A-K15 ubiquitination
Expression of isolated UBDs in HEKTLR6 cells. HEKTLR6 cells were cotransfected in triplicates using sgRNA, Cas9, pTLR-donor and unfused UBDs of Rad18 and RNF169, driven by the CAG promoter. Transfected cells were gated based on expression of BFP and the percentage of Venus (HDR) (green bars) or RFP (NHEJ) (red bars) positive cells was determined by FACS analysis. The expression of these UBDs decreases NHEJ as detectable by RFP expression but do not enhance HDR. Gene editing efficiency is expressed as percentage of Venus (HDR) (green bars) or RFP (NHEJ) (red bars) positive cells normalized to values of sample 1 (sgRosa/Cas9). Data are shown as mean values ± SD from two independent experiments, each with three replicates per samples, normalized to the values of the first (control) sample. Significance of values in comparison to the control with sgRosa/Cas9 and TLR-donor was determined by two-way ANOVA and Dunnett’s multiple comparison tests with **P < 0.01, ***P < 0.001. Raw data are shown in the Supplementary data file