Type I interferons (IFN) plays a major role in the first line of defense against viral infection. Viral RNA triggers the cytoplasmic viral sensor retinoic acid-inducible gene I (RIG-I), one of the major Pattern Recognition Receptors (PRRs), and ultimately leads to the production of type I IFN as well as pro-inflammatory cytokines through NF-kB signaling. TBK1 is a critical kinase implicated in RIG-I dependent IFN transcription. The tumor necrosis factor receptor-associated factor (TRAF) protein family members are cytoplasmic regulatory molecules that function as signal transducers for receptors in both innate and adaptive immune responses. TRAF7 belongs to the TRAF protein family and processes E3 ubiquitin ligase activity. Given that the role of TRAF7 in the regulation of type I IFN has never been investigated; and that TRAF7 has been reported to negatively regulate the NF-kB pathway, we investigated the role of TRAF7 in the regulation of type I IFN signaling. Using reporter gene assay, we demonstrated that co-expression of TRAF7 inhibits Sendai virus-mediated activation of both of the ISRE-Luc reporter and IFN-β-Luc reporter activities. Overexpression of TRAF7 in A549 cells inhibited the virus-induced endogenous IRF3 phosphorylation and ISG (IFNB1, IFIT1, and ISG15) mRNA expression. Using CRISPR/Cas9 technology, we generated TRAF7-deficient cells. Loss of TRAF7 expression in A549 cells dramatically reduced VSV infection/replication and enhanced the expression of several interferon-regulated genes. Mechanistically, TRAF7 interacts with TBK1 and promotes its degradation. We are currently generating TRAF7-deficient mice for investigating the physiological function of TRAF7 in vivo. This study reveals a previously unappreciated role for TRAF7 in the regulation of innate antiviral response.