Fig. 3. Therapeutic and multiplex base editing in β-thalassemia patient CD34⁺ HSPCs.

a Base editing by ABE8e at +58 BCL11A enhancer with sgRNA-1620 (top two rows) and HBG promoter by sgHBGsense (bottom two rows) after single or multiplex editing in β-thalassemia donor (β⁰β⁺ _#1 patient). b β-like globin expression by RT-qPCR normalized by α-globin, measured from edited erythroid progeny. Data are plotted as mean ± sd. n = 3 replicates from independent differentiation cultures. c Base editing in unfractionated BM after 16 weeks as compared to input HSPCs. Data are plotted as mean ± sd. n = 6 primary recipients for each group of engrafted HSPCs. d Human BM chimerism analyzation 16 weeks after base edited HSPC infusion. Data are plotted as mean ± sd. n = 5 mice for mock and n = 6 mice for edited group. e Percentage of engrafted human B cells, myeloid cells and CD19⁻CD33⁻ cells 16 weeks after transplantation (donor cells from β⁰β⁺ _#1 patient). Data were presented as mean ± sd, n = 5 mice for mock and n = 6 mice for edited group. f β-like globin expression analyzed by RT-qPCR normalized by α globin, measured from β⁰β⁺ _#1 patient donor BM chimerism 16 weeks after base edited HSPC infusion. Data are plotted as mean ± sd and analyzed with the unpaired two-tailed Student’s t test, p values have been noted on the corresponding comparison. n = 9 replicates for mock, and n = 11 for edited group.