Journal
ANTIVIRAL RESEARCH
Volume 90, Issue 3, Pages 187-194Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.antiviral.2011.03.174
Keywords
Gene silencing; Target control; DsiRNA; Rhabdovirus; VHSV; IHNV
Categories
Funding
- ADL-diagnostics and Chilean government [09PTEC-5238 (209-5238)]
- Danish Technical Research Council [26-03-0059]
- EEC Sixth Framework Programme [007103]
- EU Network of Excellence
- EPIZONE [FOOD-CT-2006-016236]
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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. (C) 2011 Elsevier B.V. All rights reserved.
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