Extended Data Fig. 1: Highly efficient editing of HBG promoter in CD34+ HSPCs.
a. Heat map showing the pearson correlation of mock and ABE8e edited cells transcriptomes as measured by RNA-seq. The number in each cell represents the correlation coefficient. b. Flow cytometry plots showing the expression of the erythroid maturation markers CD71 and CD235a in vitro differentiation of HUDEP-2 cells. c. Flow cytometry plots showing the expression of the RBCs maturation markers CD71 and CD235a after in vitro differentiation of ABE8e RNP edited CD34+ cells. d. Enucleation rates in erythroid cells following in vitro differentiation of ABE8e RNP edited and un-edited CD34+ cells. e. Base-editing rates in bulk base-edited cells in high and low HbF cells after in vitro differentiation of ABE8e RNP edited CD34+ cells. The T-C conversion rate of sgRNA-25 in CD34+ cells was measured by deep sequencing and quantified with CRISPResso2. f. Representative RP-HPLC chromatograms of erythroid cells derived from in vitro differentiation of edited CD34+ HSPCs. Protein levels of β-like globin by genome editing mediated by ABE8e/BCL11A Enh, ABE8e/sgRNA-25, ABE8e/sgRNA-35 and ABE8e/sgRNA-41 were analyzed. Three independent experiments were performed. Data are plotted as the mean ± s.d.
