Fig. 3

SCR7 promoted insertion repair efficiency at AAVS1 locus. a schematically illustrates the insertion repair mediated by ssODN and CRISPR/Cas9 for AAVS1, and the three methods used for the detection of insertion repair efficiency. A pair of primer P2 and P4 were used to examine insertion repair occurred in the Cas9-targeted locus by semi-quantitative PCR-gel electrophoresis. The two pairs of primers F3 and R3, F1 and R1 were used to amplify the sequences involved in the targeted site. The PCR-amplified products were analyzed by RFLP assay following EcoRI digestion and by TA cloning and DNA sequencing. b and c show representative images of PCR amplification with P2 and P4 primer and gel electrophoresis from at least three independent experiments. The data shows the enhancement of insertion repair by ssODN and SCR7 treatment in MCF-7 and HCT-116 cells in a SCR7 dose-dependent manner. “M”—DNA markers, “BC”—blank control without cells, “Con”—control cells without transfection of ssODN and CRISPR/Cas9 vector. d and e show the quantitative data of PCR-gel electrophoresis analysis using Image J software. GAPDH was used as an internal control. The data is the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01 compared to the corresponding vehicle control