Figure 4
From: A CRISPR-based approach for targeted DNA demethylation
Assessment of the off-target effect of the dCas9-based system. (a) The top 10 potential off-target sites predicted for sgRANKL-8 (R8) were selected using a scoring system described previously [16]. Sequences in the first column showed DNA sequences of the potential off-target sites; the numbers of mismatches indicated the numbers of basepairs (bp) in off-target sites that were different from those in the authentic R8-targeting site. The bp numbers indicated the locations of off-target sites in related chromosomes. (b) Changes in expression levels of RANKL mRNA in cells co-transfected with R8 sgRNA and dCas9- and MS2-Tet1-CD, compared with those of the blank or sgRNA controls. Quantitative real-time PCR (qRT-PCR) assays were performed with cells of the indicated groups. Sequences of the specified primers were listed in Supplementary Table S5. Data were shown after normalization to the controls (blank group) (means±s.e.m., n=3).
