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Infection with the nervous necrosis virus (NNV) causes viral nervous necrosis, resulting seriouslly economic losses in the aquaculture of marine fish. The viral genome of orange-spotted grouper nervous necrosis virus (OSGNNV) consists of two single-stranded, RNA1 and RNA2, encoding RNA-dependent RNA polymerase (RdRp) and capsid protein (CP), respectively. RNA interference has been shown to have activities against various viruses and is considered a promising antiviral approach. Here, we describe antiviral activities of short hairpin RNAs (shRNAs) that target RdRp and CP gene of OSGNNV in GF-1 cells. We constructed shRNAs for two specific target genes, RdRp and CP genes of OSGNNV. The shRNAs were transfected into GF-1 cells, followed by OSGNNV infection at multiplicity of infection (M.O.I) of 10.0. Then, the efficient inhibition of OSGNNV replication was examined by real-time quantitative RT-PCR analysis of RdRp and CP gene expression and viral titers. The results showed that the expression levels of RdRp and CP gene were reduced by shRNAs-transfected GF-1 cells at 48 and 72-hour post infection (hpi). Besides, the viral titers of shRNAs-transfected GF-1 cells were significantly lower than those of OSGNNV control at period of 48-96 hpi. These results indicated that the plasmid-transcribed shRNAs could inhibit effectively the OSGNNV replication in GF-1 cells, providing a potential approach to control viral disease in aquaculture. [ABSTRACT FROM AUTHOR]