Extended Data Fig. 8: Targeting erythroid-specific regulatory elements to increase HbF levels in erythroid progeny derived from HSPCs from donors with SCD.
From: Single-nucleotide-level mapping of DNA regulatory elements that control fetal hemoglobin expression
a, A heatmap showing the distribution of chromatin accessibility, as measured by ATAC-seq, near edited adenines for 15 different blood cell types. Representative adenines with high (top) and low (bottom) erythroid-specific scores (Z-scores) were selected for plotting. The cell types for each track are shown at the bottom. b–e, CD34+ HSPCs from two donors with SCD were transfected with RNP consisting of ABE and Chr11-1 gRNA targeting the 3′ HBG1 enhancer or a non-targeting control gRNA (Ctrl), then grown in culture under conditions that support erythroid differentiation. Hemoglobinized erythroblasts were analyzed at day 12. b, The percentage of γ-globin mRNA as determined by real-time RT-qPCR (n = 2 different SCD participants). c, Representative flow-cytometry plots showing the expression of the RBC maturation markers Band3 and CD49d (n = 2 different SCD participants). d, May–Grünwald–Giemsa–stained erythroblasts. Scale bar, 20 μM. This is representative results of 2 SCD participants. e, Images of sickled erythroid cells. Arrowheads mark cells with sickle-like morphology. This is representative results of 2 SCD participants. Original picture was visualized by phase-contrast microscopy using the IncuCyte S3 Live-Cell Analysis System (Sartorius) with a 20X objective; Size bars, 20 μM.
