BCL11A is required for normal erythropoiesis [ATAC-seq]

Genetic depletion of the transcriptional repressor BCL11A in red blood cell precursors alleviates b-hemoglobinopathies by inducing the fetal g-globin genes. However, several additional erythroid genes are regulated by BCL11A and the effects of its deficiency on erythropoiesis are insufficiently described. We discovered that Cas9 disruption of the BCL11A intron 2 erythroid enhancer in CD34+ hematopoietic stem and progenitor cells (HSPCs) according to a clinically approved strategy caused impaired expansion and increased apoptosis of erythroid precursors in vitro and in vivo. Mutant colony-forming unit erythroid (CFU-e), proerythroblasts, and basophilic erythroblasts exhibited dysregulation of 94 genes (> 2-fold change, FDR < 0.05). 25 of which are likely direct targets of BCL11A. Differentially expressed genes were associated with numerous biological pathways that impact cell expansion and survival. Our findings show that BCL11A regulates additional aspects of erythropoiesis beyond g-globin gene repression. ATAC-seq experiments were performed inCD34+ derived erythroidblast cells treated with CD33 sgRNA and BCL11A enhancer sgRNA