Figure 3.

Base editing mutagenesis of the IFN-γ pathway

(A) Schematic of the key mediators of IFN-γ signaling investigated in base editing screens. Depicted are top hits from our CRISPR-Cas9 screens to determine the modulators of sensitivity to IFN-γ; positive mediators are in blue and negative regulators are in red. The number of predicted LOF and GOF missense variants revealed from all base editing screens are indicated.

(B) Base editor mutagenesis of core IFN-γ pathway components using HT-29 iBE3 cells reveals GOF and LOF missense mutations. The average FACS screen score is plotted against the average proliferation screen score for each gene. Positions of validated JAK1 gRNAs and amino acid positions with predicted missense LOF or GOF effect are labeled.

(C) Base editing reveals the position of functional domains. Schematics of the domain architecture of proteins in the IFN-γ pathway tiled with base editing gRNAs, with the distribution of GOF and LOF amino acid positions labeled. All data are representative of two independent screens performed on separate days. See also Figure S3 and Table S2.