Figure 2.

PD-1-targeting CRISPR-Cas9 for TIL-based ACT is highly efficient

(A) CRISPR-Cas9-mediated PD-1 knockout throughout and post-REP measured via flow cytometry. Knockout calculated compared with Mock. See also Figure S2. (B) Surface expression of PD-1 on CD3+ TILs from mock and edited samples throughout the REP process, measured via flow cytometry. Statistical significance calculated by repeated-measures two-way ANOVA with Geisser-Greenhouse correction and Holm-Sidak multiple comparisons test (∗p ≤ 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001). See also Figure S2. (C) Representative histogram of PD-1 knockout measured by flow cytometry in post-REP stimulated sample. Mock sample shown as light gray and edited as black. (D) Representative REP day 14 IDAA plots for mock (top) and edited (bottom) samples. Yellow peaks denote unedited wild-type amplicons, blue peaks indicate frameshift indels, and white peaks indicate in-frame indels. (E) Correlation analysis of day 14 stimulated REP-TIL surface PD-1 knockout calculated via flow cytometry (compared with mock) and REP day 14 IDAA indel quantification. Correlation calculated via linear regression. Each point represents the average of two replicates for each of the 10 samples. (A and B) Each point represents the average of two replicates per sample. Bars signify median of 10 samples.