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.
Dang
Lua T
LT
Laboratory of Genome Science, Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Minato, Tokyo 108-8477, Japan.
Kondo
Hidehiro
H
Hirono
Ikuo
I
Aoki
Takashi
T
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