title: Species specific inhibition of viral replication using dicer substrate siRNAs (DsiRNAs) targeting the viral nucleoprotein of the fish pathogenic rhabdovirus viral hemorrhagic septicemia virus (VHSV).
abstract: Gene knock down by the use of small interfering RNAs (siRNAs) is widely used as a method for reducing the expression of specific genes in eukaryotic cells via the RNA interference pathway. But, the effectivity of siRNA induced gene knock down in cells from fish has in several studies been questioned and the specificity seems to be a general problem in cells originating from both lower and higher vertebrates. Here we show that we are able to reduce the level of viral gene expression and replication specifically in fish cells in vitro. We do so by using 27/25-mer DsiRNAs acting as substrates for dicer for the generation of siRNAs targeting the nucleoprotein N gene of viral hemorrhagic septicemia virus (VHSV). This rhabdovirus infects salmonid fish and is responsible for large yearly losses in aquaculture production. Specificity of the DsiRNA is assured in two ways: first, by using the conventional method of testing a control DsiRNA which should not target the gene of interest. Second, by assuring that replication of a heterologous virus of the same genus as the target virus was not inhibited by the DsiRNA. Target controls are, as we have previously highlighted, essential for verification of the specificity of siRNA-induced interference with virus multiplication, but they are still not in general use.
authors: ADL Diagnostic Chile Ltda, Laboratorio de Diagnóstico y Biotechnología, Puerto Mont, Chile.
authors: Harry
authors: H
authors: Bohle
authors: Niels
authors: N
authors: Lorenzen
authors: Brian Dall
authors: BD
authors: Schyth
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D015536:Down-Regulation; D031941:Hemorrhagic Septicemia, Viral / Q000201:enzymology / Q000628:therapy* / Q000821:virology*; D008969:Molecular Sequence Data; D029141:Novirhabdovirus / Q000235:genetics* / Q000502:physiology; D019590:Nucleocapsid Proteins / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism / Q000627:therapeutic use; D043244:Ribonuclease III / Q000378:metabolism*; D012474:Salmon; D013045:Species Specificity; D014779:Virus Replication / Q000187:drug effects
chemical_list: D019590:Nucleocapsid Proteins; D034741:RNA, Small Interfering; D043244:Ribonuclease III