Fig. 2: A genome-wide CRISPR screen identifies the HGS and VPS37A ESCRT subunits as required for STING degradation.
From: ESCRT-dependent STING degradation inhibits steady-state and cGAMP-induced signalling
a mNeonGreen (mNG) signal intensity in 293T (gray) or a 293T cell line stably expressing a STING-mNG reporter non-stimulated (NS) (green) or stimulated (red) with 1 µg/ml 2′3′-cGAMP(pS)2 (in medium) for 24 h. One representative experiment of n = 3 experiments. b Strategy for genome-wide CRISPR screen to identify regulators of STING trafficking and degradation. c Volcano-plot of log2 fold change (log2FC) vs -log10(pvalue) after sequencing and analysis of the genome-wide CRISPR screen as in b). Genes of interest are highlighted in red. Control guides are in blue. d Percentage of STING-mNG positive 293T STING-mNG in cells stimulated or not with 1 µg/ml 2′3′-cGAMP(pS)2 (in medium) for 24 h. Shown is ratio %STING-mNG positive of each sgRNA over %STING-mNG positive cells of the control non-targeting sgRNA (ntgRNA). Two independent sgRNAs per gene. n = 2 independent experiments with n = 2 technical replicates per experiment. Each dot represents an individual replicate. One-way ANOVA with Dunnet’s multiple comparisons test. ****p < 0.0001, ***p < 0.001, *p < 0.05, ns: not-significant. e Intersection of all proteins identified by STING-TurboID proteomics (TurboID all) and filtered proteins (TurboID filtered) with hits from the genome-wide CRISPR screen with log2FC > 0 and −log10(pvalue)>2.
