Extended Data Fig. 5: Optimization and characterization of B2M targeted SEEDs.
From: SEED-Selection enables high-efficiency enrichment of primary T cells edited at multiple loci
a–e, T cells were edited with RNP targeting B2M and transduced with varying MOIs of AAV encoding CD47. In indicated conditions, editing was performed in the presence of M3814 (n = 3 donors). Cells transduced at an MOI of 1 × 104 and edited in the presence of M3814 shown in select panels (a,b). a, Representative flow cytometry plots of B2M and CD47 expression in edited and non-edited cells. b, Median fluorescence intensity (MFI) of B2M or CD47 expression in subpopulations of edited cells. Data displayed as mean plus SEM. Significance was assessed using a repeated-measures one-way ANOVA and Turkey’s multiple comparison test. c, Percentage of CD47+ cells, d, Percentage of B2M– CD47+ cells, e, Percentage of CD47+ cells with full B2M disruption. Data for c–e displayed as mean plus SEM. f,g, T cells edited with RNP targeting B2M ( + M3814) and transduced with high (3 × 105) or low (1 × 104) MOIs of B2M-CD47 SEED HDRT AAV were immunomagnetically purified with anti-B2M (n = 3 donors). f, Representative flow cytometry plots of B2M and CD47 expression. g, Percentage of B2M– CD47+ cells. Data displayed as mean plus SEM (n = 3 donors). h, Representative flow cytometry plots of B2M and CD47 expression used to evaluate CD47 + :CD47 ratio in Fig. 1h.
