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 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 36h 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.
Yin
Renfu
R
Department of Veterinary Preventive Medicine, College of Animal Science and Veterinary Medicine, Jilin University, Xi'an Road 5333, Changchun, Jilin 130062, China.
Ding
Zhuang
Z
Liu
Xinxin
X
Mu
Lianzhi
L
Cong
Yanlong
Y
Stoeger
Tobias
T
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002478:Cells, Cultured; D002642:Chick Embryo; D005347:Fibroblasts / Q000821:virology*; D009522:Newcastle disease virus / Q000235:genetics / Q000254:growth & development*; D010957:Plasmids; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000494:pharmacology*; D019562:Viral Load; D014763:Viral Matrix Proteins / Q000037:antagonists & inhibitors*
D000998:Antiviral Agents; D034741:RNA, Small Interfering; D014763:Viral Matrix Proteins