Figure 10.
The influence of MBD4 knockdown on the efficiency of gene editing by a methyl-CpG-modified ssODN. (A) Expression of GFP protein was followed over time by FACS analysis. Myoblast cultures stably expressing the mutant GFP were co-transfected with m5CpGGFP ssODN and either a control siRNA or an siRNA capable of downregulating MBD4 expression (or with no siRNA). The percentage of GFP-positive cells was significantly decreased 24 and 48 h after transfection in cultures treated with the targeting siRNA compared with the control siRNA or no siRNA. *P ≤ 0.002. (B) Real-time PCR was used to determine gene editing efficiencies of cells transfected with the m5CpG ssODN and with either a control siRNA or the MBD4 siRNA. Genomic DNA was isolated 24 and 48 h after transfection and digested with BtsI. The levels of the amplification products in cultures treated with the control siRNA were set at 100%. Amplification products were significantly reduced in cells treated with the MBD4 siRNA at 24 h and further reduced at 48 h. *P ≤ 0.02; **P ≤ 0.004.