Functional analysis of wssv025 gene in the pathogenicity of white spot syndrome virus in Penaeus monodon

White Spot Syndrome Virus (WSSV) is an extremely virulent pathogen that causes significant financial damage in shrimp aquaculture. Despite extensive research, the role of specific viral genes such as wssv025 in pathogenesis remains unclear. In this study, wssv025 is investigated for its role in WSSV pathogenesis and potential as a target for antiviral strategies in shrimp aquaculture. The wssv025 protein interacts with several key viral proteins in yeast two-hybrid (Y2H) assays, suggesting that it plays an important role in viral replication, immune evasion, and host modulation. Silencing wssv025 significantly reduced the transcriptional and translational levels of major WSSV genes such as vp28, icp11, and ie1. The Western blot analysis confirmed these findings with wssv025 gene silencing leading to a significant reduction in viral protein expression. A tissue-specific response to wssv025 was found by analyzing the viral genome copy numbers in pleopods and viral gene expression in hemocytes. The results of this study underscore the complex relationship between viral replication, host immune responses, and gene expression. The study provides strong evidence supporting RNAi-based antiviral strategies targeting wssv025 as a potential therapeutic approach for controlling WSSV infections in giant tiger prawn (Penaeus monodon).