Inhibition of orange-spotted grouper nervous necrosis virus replication by short hairpin RNAs in grouper GF-1 cells

Document Type : Original research

Authors

• Minh Thi Tuyet Vo ¹
• Cheng-Hui Lin ²

¹ Faculty of Agriculture and Aquaculture, Tra Vinh University, Viet Nam

² Department of Aquaculture, National Taiwan Ocean University, No. 2, Pei-Ning Road, Keelung 20224, Taiwan

10.22034/iar.2022.1971400.1344

Abstract

Infection with the Nervous necrosis virus (NNV) causes viral nervous necrosis, resulting serious economic losses in the aquaculture of marine fish. The viral genome of orange-spotted grouper nervous necrosis virus (OSGNNV) is composed of two single-stranded, RNA1 and RNA2, encoding RNA-dependent RNA polymerase (RdRp) and capsid protein (CP), respectively. Small interfering RNAs, which mediate a sequence-specific gene silencing, have been demonstrated to have activity against viruses and are beeing explored as prospective antiviral tools. Here, we describe the antiviral activity of short hairpin RNAs (shRNAs) that target RdRp and CP gene of OSGNNV in grouper GF-1 cells. Inhibition of OSGNNV replication was demonstrated by reduced RdRp and CP gene expression and reduced OSGNNV titer. The expression of RdRp and CP gene was inhibited by shRNAs in GF-1 cells transfected with OSGNNV at 48 and 72-hour post infection (hpi). Furthermore, the OSGNNV titer in shRNA-transfected GF-1 cells was significantly lower than that of OSGNNV group at period of 48-96 hpi. These results indicate that the plasmid-transcribed shRNAs can efficiently inhibit the replication of OSGNNV in GF-1 cells, providing a potential approach to control viral disease in aquaculture.

Keywords

• Orange-spotted grouper nervous necrosis virus (OSGNNV)
• shRNAs
• RdRp gene expression
• CP gene expression
• Transfection
• Inhibition
• GF-1 cells

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