Inhibition of Newcastle disease virus replication by RNA interference targeting the matrix protein gene in chicken embryo fibroblasts

Newcastle disease (ND) is an infectious viral disease of birds caused by the Newcastle disease virus (NDV), also known as avian paramyxovirus type 1 (AMPV-1), which leads to severe economic losses in the poultry industry worldwide. In this study, the application of RNA interference (RNAi) for inhibi...

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Bibliographic Details
Published in:Journal of virological methods 2010-07, Vol.167 (1), p.107-111
Main Authors: Yin, Renfu, Ding, Zhuang, Liu, Xinxin, Mu, Lianzhi, Cong, Yanlong, Stoeger, Tobias
Format: Article
Language:English
Subjects:
Animals
Antiviral activity
Antiviral Agents - pharmacology
Biological and medical sciences
Cells, Cultured
Chick Embryo
Fibroblasts - virology
Fundamental and applied biological sciences. Psychology
Inhibition
Microbiology
Newcastle disease virus
Newcastle disease virus - genetics
Newcastle disease virus - growth & development
Plasmids
RNA Interference
RNA, Small Interfering - pharmacology
Short hairpin RNA
Techniques used in virology
Viral Load
Viral Matrix Proteins - antagonists & inhibitors
Virology
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Summary:Newcastle disease (ND) is an infectious viral disease of birds caused by the Newcastle disease virus (NDV), also known as avian paramyxovirus type 1 (AMPV-1), which leads to severe economic losses in the poultry industry worldwide. In this study, the application of RNA interference (RNAi) for inhibiting the replication of NDV in cell culture by targeting the viral matrix protein gene (M) is described. Two M-specific shRNA-expressing plasmid constructs, named pS M641 and pS M827, were evaluated for antiviral activity against the NDV strain NA-1 by cytopathic effects (CPE), virus titration and real-time RT-PCR. After 36 h of infection, both pS M641 and pS M827 reduced virus titers by 79.4- and 31.6-fold, respectively, and they down-regulated mRNA expression levels of the matrix protein gene M by 94.6% and 84.8%, respectively, in chicken embryo fibroblast (CEF) cells, while only pS M641 significantly decreased CPE, compared to the control group. These results indicated that the M gene 641 and 827 sites represent potential antiviral therapy targets, and RNAi targeting of the M gene could not only represent an effective treatment in Newcastle disease but also aid as a method for studying the replication of NDV.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2010.02.014