Inhibition of red seabream iridovirus (RSIV) replication by small interfering RNA (siRNA) in a cell culture system

• Published in: Antiviral research Vol. 77; no. 2; pp. 142 - 149
• Main Authors: Dang, Lua T., Kondo, Hidehiro, Hirono, Ikuo, Aoki, Takashi
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.02.2008; Elsevier
• Subjects: Animals; Antibiotics. Antiinfectious agents. Antiparasitic agents; Antiviral agents; Base Sequence; Biological and medical sciences; Capsid Proteins - genetics; Capsid Proteins - metabolism; Cell Line; Cells, Cultured; Iridovirus; Iridovirus - genetics; Major capsid protein; MCP; Medical sciences; Pharmacology. Drug treatments; Plasmids; Red seabream iridovirus; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; RNA interference (RNAi); RNA, Small Interfering - genetics; RSIV; Sea Bream - virology; Small interfering RNA (siRNA); Transfection; Virus Replication - drug effects; RSIV; RNA interference (RNAi); Small interfering RNA (siRNA); MCP; Red seabream iridovirus; Major capsid protein; Cell culture; RNA interference; Pagrus major; siRNA; In vitro; Protein; Virus; Gene silencing; Vertebrata; Iridoviridae; Pisces; Replication; Cell system; Capsid; Veterinary; Iridovirus
• ISSN: 0166-3542; 1872-9096
• DOI: 10.1016/j.antiviral.2007.10.007

Small interfering RNAs (siRNAs), mediators of a process of sequence-specific gene silencing called RNA interference, have been shown to have activity against a wide range of viruses and are considered to be potential antiviral tools. Here, we describe an antiviral activity of a siRNA that targets the major capsid protein (MCP) gene of red seabream iridovirus (RSIV), a marine fish-pathogenic virus, in a cell culture system. Inhibition of RSIV replication was demonstrated by reduced MCP expression level and reduced RSIV titer. MCP-targeted siRNA (siR-MCP) dose-dependently inhibited the expression of MCP gene in cells that either transiently expressed or stably expressed the MCP gene. At 84 and 96 h after viral infection, siR-MCP reduced the expression of MCP gene by 55.2% and 97.1%, respectively. Transfection with siR-MCP reduced the production of RSIV particles in supernatants of samples infected with RSIV, while the corresponding mismatched siR-MCP (MsiR-MCP) and nsRNA controls did not exhibit this effect. These results show that MCP-targeted siRNA can effectively and specifically inhibit the expression of the target gene and hinder RSIV replication during an in vitro infection, providing a potential approach for the control of viral diseases in aquaculture.