Screening and Validation of Negative Regulators of the Type I Interferon Pathway

Abstract Type I interferons （IFN‐I） are key proteins in antiviral response and immunomodulation. Negative regulators avoid abnormal activation of the interferon pathway or overactivation of interferon‐activating proteins through multiple mechanisms. Loss‐of‐function mutations in negative regulator genes lead to the development of a variety of type I interferonopathy. It is of great significance in theory and clinic to discover new negative regulators and study their functions, but there is no effective screening system at present. Here, a screening system is established through a resistance reporter, which is designed to be expressed only in cells with interferon pathway activation. In conjunction with CRISPR knockout library, cells are screened for interferon pathway activation due to knockout of negatively regulators, which led to the identification of negatively regulator candidate genes such as PCGF3/5 , UCK2 , and ITPKA . The expression products of these genes functioned by targeting MAVS and promoted EMCV (encephalomyocarditis virus) infection by inhibiting interferon pathway activation. This study deepens the understanding of the regulatory network of the interferon pathway and provides a new theoretical basis for the study of the pathogenesis of autoimmune diseases.