Reactivation of Zeta-Globin by deletion of BCL11A and LRF in HUDEP cells (ATAC-seq)

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. Overall design: CRISPR/Cas9 editing was carried out to generate homozygous clones where LRF, BCL11A and BCL11A/LRF were homozygously deleted. Effects of edited variants were determined by ATAC-seq in technical replicates. Cells were sorted to obtain normoblasts and ensure homogeneity (strong CD235a, weak CD49d, strong Band3). Experiments were carried out at Day 10 of differentiation.