Expression data from HEK293T cells

Abnormal DNA sensing mediated by cGAS-STING is pivotal for host defenses. Activation of cGAS-STING triggered interferon dependent anti-viral functions have been well characterized but this function is missing in more ancient animal STING. Whether more primitive animal STING can generate non-canonical pathway dependent anti-viral activities is unclear. Here, we discovered potent anti-viral activities of ancient animal STING in the absence of all three canonical pathways. Various ancient animal STING-stimulated genes (ASGs) with unique and synergistic anti-viral functions were induced by this newly identified pathway through an ancient transcriptional regulator IRF1 and a separate unique functional domain in STING. Some ASGs are both necessary and sufficient as host restriction factors and under positive selection during animal evolution. Diverse viruses have developed strategies to evade this newly identified cGAS-STING function. Our findings reveal an ancient, conserved cGAS-STING pathway mediated anti-viral activities through previously unrecognized cellular restriction factors. Various animal STING expression in HEK293T cells could up regulate a series of genes. To screen candidates for anti-viral host factors stimulated by STING in the absence of the known canonical pathways, we identified the factors that were regulated by the IFN induction and NF-κB induction-defective Nematostella vectensis (N.v) STING and the IFN induction-defective human STING variant (H.s STING ΔCTT) by microarray assay. Multiple rounds of confirmative and comparative experiments narrowed down to 10 such factors, i.e., ancient animal STING stimulated genes (ASGs). ASGs were significantly upregulated by STING activation in various human cell types. The H.sSTING ΔCTT, N.vSTING or empty vector were co-transfected with cGAS in HEK293T cells. Twenty hours later, mRNA samples were prepared and analyzed using Affymetrix microarrays. We sought to obtain up-regulated genes by the IFN induction-defective human STING variant (H.s STING ΔCTT) and IFN induction and NF-κB induction-defective N.v STING