Inhibition of red seabream iridovirus (RSIV) replication by small interfering RNA (siRNA) in a cell culture system.
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 96h 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.
Laboratory of Genome Science, Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Minato, Tokyo 108-8477, Japan.
Lua T
LT
Dang
Hidehiro
H
Kondo
Ikuo
I
Hirono
Takashi
T
Aoki
D000818:Animals; D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D017994:Iridovirus / Q000235:genetics*; D010957:Plasmids; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D021541:Sea Bream / Q000821:virology; D014162:Transfection; D014779:Virus Replication / Q000187:drug effects*
D036022:Capsid Proteins; D034741:RNA, Small Interfering; C081132:major capsid protein, Iridovirus