Janus Kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathway plays an important role for antiviral immunity. In brief, the signal binding to DOME receptor activates JAK, leading to phosphorylated STAT and their dimerization. STATs, then, localized to the nucleus and activated gene transcription. In order to better understand the role of JAK/STAT signaling pathway in Penaeus monodon during viral infection, RNA interference technique was employed to silence the expression of PmDOME and PmSTAT gene expression in Penaeus monodon, and the cumulative mortality of shrimp were observed after WSSV infection. From tissue distribution, PmDOME, PmJAK and PmSTAT were expressed in all tested tissues. Knockdown of PmDOME and PmSTAT delayed shrimp mortality after WSSV challenge. In addition, PmSTAT silencing has altered some gene expression in the immune deficiency (Imd) pathway and antimicrobial peptides (AMPs).