Fig. 5: HBG promoter demethylation activates gene expression in erythroblasts derived from CD34+ cells.
From: Removal of promoter CpG methylation by epigenome editing reverses HBG silencing
Healthy donor CD34+ cells were electroporated with TETv4 mRNA or dTETv4 mRNA + six sgRNAs targeting the HBG promoter or sgRNA targeting a control gene (EPX), followed by in vitro erythroid differentiation. a Bisulfite Amplicon sequencing at day 12 of differentiation showing methylation levels at the indicated CpG sites flanking the HBG promoter. Data show mean percentage ± s.d. of three biological replicates. b %HBG mRNA following induced erythroid differentiation. Data show mean percentage ±s.d. of three biological replicates. c %HbF determined by ion exchange HPLC following induced differentiation. Data show mean percentage ± s.d. of three biological replicates. d Representative FACS profiles of erythroid maturation markers CD235a, CD49d, and Band3 at day 15 of differentiation. e Epigenetic analysis of the extended β-like globin locus in TETv4-treated or control dTETv4-treated erythroblasts on day 10 of erythroid differentiation. ATAC-seq analysis indicates chromatin accessibility. CUT&RUN analysis shows the occupancy of GATA1, NFYA, and BCL11A, and H3K4me3 modifications. ****P < 0.0001, Multiplicity-adjusted P values by one-way analysis of variance (ANOVA) for (b, c). Source data are provided as a Source Data file.
