Analysis of nascent transcription by PRO-seq in HUDEP-2 cells following CRISPR editing of BCL11A enhancer

CRISPR editing of an intronic enhancer within the BCL11A gene reactivates fetal hemoglobin (HbF) in adult erythroid cells, serving as a gene-based therapy for hemoglobinopathies. However, the mechanisms underlying the remarkable efficacy of CRISPR-mediated enhancer ablation remain poorly understood. Ehancer RNAs transcribes from enhancer play a role in gene regulation. Here, we apply PRO-seq in wild-type and sg1617-edited HUDEP-2 cells to determine the effects of sg1617 editing on enhancer RNA transcription. Overall design: PRO-seq was performed in wild-type HUDEP-2 cells and sg1617-edited single-cell clones with 1 bp insertion, 13 bp deletion, or 15 bp deletion.