Ubiquitin-binding enzyme E2J1 (UBE2J1) is a predominant member of the UBE2 family. In mammals, it is involved in antiviral immunity, ER-misfolded protein degradation, and ubiquitination. However, the function of UBE2J1 in teleost has not been elucidated. In the present study, we cloned the homolog of UBE2J1 in triploid hybrid fish and investigated its regulatory role in the antiviral interferon (IFN) signaling. We determined that the full-length coding sequence of the triploid fish UBE2J1 (3nUBE2J1) consists of 939 base pairs and encodes 313 amino acids. Upon SVCV infection, the mRNA levels of 3nUBE2J1 were increased in the spleen, liver, kidney, and the caudal fin cell line of triploid fish. Knockdown of 3nUBE2J1 in host cells and fish enhanced their antiviral response and curtailed SVCV replication. Conversely, overexpression of 3nUBE2J1 inhibited the activation of IFN promoter induced by SVCV, poly (I:C), or RLR pathway factors. Further studies revealed that 3nUBE2J1 interacted with 3nIRF7 and suppressed both the protein level and antiviral activity of 3nIRF7. Finally, we demonstrated that 3nUBE2J1 enhanced the K27- and K29-linked ubiquitination of 3nIRF7, leading to its degradation. Collectively, our findings indicate that triploid fish UBE2J1 serves as a novel negative regulator of IFN signaling by inhibiting 3nIRF7.