Skip to main content
. 2022 May 20;13:2833. doi: 10.1038/s41467-022-30465-7

Fig. 3. ABR-001 enables ex vivo editing of human primary cells.

Fig. 3

a, b Indel activity (a) and viability (b) of human CD3 + T cells following delivery of ABR-001 RNPs with 4 different B2M-targeting guides (B2M 1–4) at decreasing concentrations (16 µM, 10 µM, 5 µM, and 2 µM). Cell viability was measured using flow cytometry at 7 days post-electroporation. Bars represent the average of 3 donors (circles), with error bars representing the s.d.. c Representative histograms show B2M protein level knockdown at 7 days post-electroporation with ABR-001 RNP with B2M 1 guide at decreasing concentrations (16 µM, 10 µM, 5 µM, and 2 µM). Y-axes represent count normalized to mode. d Indel activity of CD3+ T cells following delivery of 16 µM ABR-001 RNPs targeting TRAC and CIITA. Circles represent each of n = 2 bioreplicates. e Viability of CD3+ T cells following delivery of 16 µM ABR-001 RNPs targeting TRAC and CIITA. Circles represent each of n = 2 bioreplicates. f Representative histograms show protein level knockdown at 7 days post-electroporation for TRAC and CIITA (HLA-DR/DP/DQ readout). Y-axes represent count normalized to mode. g Indel size was compared between two independent experiments (BR1 and BR2) of ABR-001 RNPs targeting TRAC and CIITA. The color scale indicates the fraction of indel reads at a given indel size. h Indel rates (teal) and % GATAA disruption (black) of multiplexed ABR-001 RNPs (16 µM total concentration) or single SpCas9 RNP (5 µM concentration) targeting the BCL11A enhancer, 72 h post-delivery into human CD34+ cells. Bars represent mean and error bars represent s.d. of n = 3 replicates. i Indel rates (teal) and % GATAA disruption (black) of ABR-001 RNPs (20 µM total concentration) and SpCas9 RNP (5 µM concentration) after delivery and 20 days of maturation in erythroid differentiation media. Bars represent mean and error bars represent s.d. of n = 3 replicates. j Indel profiles of ABR-001 RNP (target 2 and multiplex target1+2) and SpCas9 RNP targeting the BCL11A enhancer in human CD34+ cells. The color scale indicates the fraction of indel reads at a given indel size. k Representative flow cytometric analyses of edited CD34+ cells after delivery and 20 days of maturation in erythroid differentiation media. Source Data are provided as a Source Data file.