Fig. 1
From: Hit-and-run epigenetic editing prevents senescence entry in primary breast cells from healthy donors
dCas9 3A3L targeted hypermethylation of HIC1 and RASSF1 in primary myoepithelial cells. a Schematic of experimental design, 10 plasmids encoding gRNAs targeting HIC1 and RASSF1A promoters were transfected along with dCas9 3A3L or 3A3LΔ and the MACS (magnetic activated cell sorting) plasmid (pMACS) for magnetic selection. b A schematic depicting the gRNA location (grey arrows) in relation to the CGI (in green) and the TSS in the promoter of the HIC1 gene; the arrows point towards the PAM sequence. The dotted lines represent the region analysed by bisulfite cloning and the locations of individual CpGs are scaled according to their genomic distribution. The dark blue shading depicts which CpGs are overlapped by gRNA #5. Lower panel shows bisulfite cloning results from the HIC1 locus 5 days after transfecting primary myoepithelial cells from donor 1 with 10 gRNAs (targeting HIC1 and RASSF1) and dCas9 3A3L or 3A3LΔ as indicated. c A schematic depicting gRNA in relation to CGI and TSS in the promoter of RASSF1A. Lower panel shows bisulfite cloning from RASSF1A 5 days after transfection of primary human myoepithelial cells with 10 gRNAs (targeting HIC1 and RASSF1) and dCas9 3A3L or 3A3LΔ as indicated. Arrows and green shading represent overlap between gRNAs and CpGs. Each line represents a single clone, filled and empty lollipops represent a single methylated or unmethylated CpG, respectively
