Extended Data Fig. 8 |. Characterization of a PIK3CD allelic series.

a, Bar graphs corresponding to Fig. 3i show LFC (log2-fold change) in expression of the indicated cytokines in arrayed validation for a series of 9 PIK3CD guides relative to AAVS1 control in CD8⁺ T cells; color indicates LFC (log2-fold change) value of each guide in the original ABE TNFα screen. n = 6 human donors, shown as mean ± SE. b, A-T to G-C editing by co-electroporation of ABE mRNA and synthetic PIK3CD sgRNAs, assessed by deep amplicon sequencing and analyzed with Crispresso2⁵⁴. Guide sequences are in gray, predicted editing window in green, and PAM in dark gray. c, Correlation between PIK3CD LFC (log2-fold change) values from the TNFα screen and validation experiment (Fig. 3i) in CD4⁺ T cells. d, mRNA expression of PIK3CD and IL2RA in PIK3CD edited T cells. Mean ± SD, n = 4 independent donors. e-g, Single amino acid substitution by CRISPR/Cas9 knockin. All edits include a synonymous K282K mutation deleting the PAM site. e, Flow cytometry histograms showing TNFα expression in CD4⁺ gated T cells. Percent positive cells in gray. f, Percent cytokine positive CD4⁺ and CD8⁺ gated T cells with the indicated knockins. n = 2 human donors (red and blue dots) shown as mean. g, Corresponding gene editing outcomes for knockin experiments as reported by CRISPResso2. n = 2 human donors shown as mean.