Supplementary Figure 5: N-terminal Nuclear Localization Signal (NLS) sequences increase the efficiency and range of base editing
From: Optimized base editors enable efficient editing in cells, organoids and mice
a. Schematic showing location of NLS sequences and linker size in each construct tested. To provide a fair comparison, each of the constructs shown carries the original (non-optimized) cDNA sequence. b. Frequency (%) of C>T conversion in co-transfected HEK293T cells with BE3, 2X, FNLS, FLAGlink, or BE4 CMV vectors and either FANCF.S1 or CTNNB1.S45 sgRNAs, as indicated. Graphs show mean values. Error bars represent s.e.m., n = 2–6 biologically independent experiments, as indicated; asterisks (*) indicate a significant difference (p<0.05) between groups, using a two-way ANOVA with Tukey’s correction for multiple testing. c. Frequency (%) of C>T conversion in the last edited cytosine relative to the first edited cytosine for each construct co-transfected with either FANCF.S1 or CTNNB1.S45 sgRNAs. Graphs show mean values. Error bars represent s.e.m., n=2-6 biologically independent experiments, as indicated; first number refers to FANCF.S1, the second to CTNNB1.S45. The BE3 condition for FANCF.S1 could not be calculated for more than one replicate as the other two showed zero editing at C11. Asterisks (*) indicate a significant difference (p<0.05) between groups, using a two-way ANOVA with Tukey’s correction for multiple testing.
