Reactivation of Zeta-Globin by deletion of BCL11A and LRF in murine cells

The alpha and beta-globin loci harbour genes that are expressed during development and silenced throughout post-natal life. Learning to reactivate these genes may offer new therapeutic approaches for the hemoglobinopathies, the most common single gene disorders. Here, we address the mechanisms underlying the regulation of the gene encoding zeta-globin, the embryonically expressed alpha-like globin. To determine the role of LRF and Bcl11a in regulating zeta globin in definitive mouse erythroblasts, mouse lines lacking LRF or the BCL11A erythroid enhancer were generated. Lrf-flox (Maeda 2007; PMID: 17495164) mice were crossed to mice expressing Cre under control of Gata1 regulatory elements (Mao 1999; PMID: 10220414), leading to germline excision of floxed regions. Heterozygous Lrf-Flox/- mice were interbred to make genetic combinations. Bcll11a-EnhKO lines lack a 10kb region which contains the mouse erythroid Bcl11a enhancer (Smith 2016; PMID: 27707736); genetic combinations were generated by crossing. Effects of edited variants were determined by ATAC-seq in technical replicates.