Fig. 1
Assessment of guide RNA activity for gene disruption at PDCD1, TRAC, and B2M. a Diagram of PDCD1 locus indicating the relative locations of each sgRNA. Colored portion of boxes represent protein-coding region, vertical red line indicates stop codon. b Quantification of C to T conversion of target base for each PDCD1 sgRNA (n = 3 independent T-cell donors). c PDCD1 protein knockout frequency (n = 3 independent T-cell donors). d Quantification of C to T/A/G conversion at all Cs within the detected editing window (shown in red) of the PDCD1 Ex1 SD sgRNA (n = 3 independent T-cell donors). Underlined C indicates target nucleotide critical for proper splicing. e Diagram of TRAC locus indicating the relative locations of each sgRNA. f Quantification of C to T conversion at target base for each TRAC sgRNA (n = 3 independent T-cell donors). g TRAC protein knockout frequency as determined by flow cytometry for CD3 loss (n = 3 independent T-cell donors). h Quantification of C to T/A/G conversion at all cytosines within the detected editing window (shown in red) of the TRAC Ex3 SA sgRNA (n = 3 independent T-cell donors). i Diagram of B2M locus indicating the relative locations of each sgRNA. j Quantification of C to T conversion of target base for each B2M sgRNA (n = 3 independent T-cell donors). k B2M protein knockout frequency (n = 3 independent T-cell donors). l Quantification of C to T/A/G conversion at all cytosines within the detected editing window (shown in red) of the B2M Ex1 SD sgRNA (data represented as mean ± SD, n = 3 independent biological T-cell donors). P-values calculated by the Student’s paired two-tailed t test between the highest-editing guide and the second highest-editing treatment (n.s. P > 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001)
