Beta-hemoglobinopathies are severe genetic diseases caused by mutations in the β-globin locus. One option for novel treatment involves the transplantation of autologous, genetically corrected HSCs. Genome editing of the BCL11A erythroid-specific enhancers using site-specific nucleases shows efficient reactivation of foetal hemoglobin (HbF) expression. Yet, HSCs are highly sensitive to DNA damage, which might result in unpredictable genotoxic effects. The recent adoption of designer epigenome modifiers (DEMs) to deposit repressive marks in a targeted manner offers the opportunity to achieve erythroid-specific BCL11A inactivation without altering the underlying genomic sequence. We anticipate that epigenetic inactivation of these BCL11A enhancers will counteract the developmental silencing of HbF and rescue the hematologic and pathologic features of β-hemoglobinopathies. We characterised the epigenetic marks decorating the erythroid-specific BCL11A enhancers during HSC differentiation toward the erythroid lineage using an established in vitro assay, and identified regions that differentially lose DNA methylation leading to enhancer activation. We produced DEMs targeting these regions to induce persistent inactivation of the erythroid-specific BCL11A enhancers. We will deliver in vitro transcribed mRNA encoding for selected DEMs into HSCs via nucleofection and will correlate the effects of epigenome editing with transcriptional changes of BCL11A and β-like globin genes. Furthermore, we will assess whether this approach leads to production of HbF and HbF+ cells after in vitro differentiation of epigenetically edited HSCs. We believe that this study will be instrumental to understand the key epigenetic features that control the γ-globin switch and will provide a novel therapeutic approach for patients with β-hemoglobinopathies.