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Chibby, a vital inhibitor molecule of Wnt/β-catenin signaling pathway, participates in development and stem cell differentiation through the regulation of β-catenin. Our previous studies have demonstrated that Litopenaeus vannamei β-catenin (Lv-β-catenin) was involved in WSSV infection and could inhibit virus replication by modulating the host immune system. In the study, a Chibby homolog from L. vannamei (designed as Lv-Chibby) was isolated and its role in WSSV infection was investigated. Sequence analysis suggested that Lv-Chibby was a novel homolog of Chibby family. It could transcript in all examined tissues, including hemocyte, gill, intestine, hepatopancreas, muscle and heart. Real-time quantitative PCR demonstrated that Lv-Chibby could take part in WSSV infection and be down-regulated by WSSV. Further studies confirmed that Lv-Chibby was able to interact with Lv-β-catenin. Moreover, the relationship of Lv-β-catenin, Lv-Chibby and WSSV069 was investigated. It was shown that Lv-Chibby enhanced the interaction between Lv-β-catenin and WSSV069. Interestingly, WSSV069 promoted the interaction between Lv-β-catenin and Lv-Chibby under high concentration, while low concentration of WSSV069 inhibited their interaction. A subsequent immunofluorescence assay revealed that WSSV069 appeared to reduce the nuclear entry of Lv-β-catenin. In sum, these results implied that Wnt/β-catenin signal pathway plays an important role in the defense against virus, and Chibby could be modulated by WSSV to regulate the signal pathway.A novel Chibby homolog from Litopenaeus vannamei was characterized.Lv-Chibby was associated with WSSV infection.Lv-Chibby and WSSV069 could regulate the interaction with β-catenin.WSSV069 reduced the nuclear entry of Lv-β-catenin.