Extended Data Fig. 2: Erythroid differentiation of CD34+ HSPCs after editing with ABE7.10 or Cas9 nuclease.
From: Potent and uniform fetal hemoglobin induction via base editing
Erythroid differentiation of CD34+ HSPCs after editing with ABE7.10 complexed with sgRNA −175 or Cas9 nuclease complexed with sgRNA targeting the BCL11A binding site in the γ-globin promoter, the +58 BCL11A erythroid enhancer or the control locus AAVS1. Cells were edited by electroporation with RNPs, incubated in CD34+ expansion medium for two days, then transferred to erythroid differentiation medium. a, Cell expansion after base editing (n = 2 from two donors). b, Cell expansion after Cas9 editing (n = 3 from one donor). c, Representative flow cytometry scatter plot showing maturation markers in CD235a+ erythroblasts after 7 and 14 days of in vitro erythroid differentiation. d, Summary of multiple experiments to assess cell maturation using gating strategy depicted in panel c. n = 2 replicates each from two donors for ABE7.10 and n = 3 from one donor for Cas9. e, Representative flow cytometry scatter plot showing enucleated reticulocytes distinguished by loss of staining with the DNA-binding dye Hoechst 33342 (gated). f, Percentage of enucleated CD235a+ erythroid cells (reticulocytes) at differentiation day 21 (n = 4 from two donors for ABE7.10 and n = 3 from one donor for Cas9). Graphs show mean ± SD. UT, untreated. NT, non-targeting gRNA.
