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Effective inhibition of infectious bursal disease virus replication in vitro by DNA vector-based RNA interference

文献类型： 外文期刊

作者： Gao, Yulong 1 ; Liu, Wei 1 ; Gao, Honglei 1 ; Qi, Xiaole 1 ; Lin, Huan 1 ; Wang, Xiaomei 1 ; Shen, Rongxian 1 ;

作者机构： 1.Chinese Acad Agr Sci, Harbin Vet Res Inst, Div Avian Infect Dis, Natl Key Lab Vet Biotechnol, Harbin 150001, Peoples R China

2.Heilongjiang Acad Agr Sci, Postdoctoral Workstn, Harbin 150086, Peoples R China

3.Heilongjiang Acad Agr Sci, Postdoctoral Workstn, Harbin 15008

关键词： RNAi;siRNA;shRNA;infectious bursal disease virus (IBDV);virus replication

期刊名称：ANTIVIRAL RESEARCH （ 影响因子：5.97； 五年影响因子：5.801 ）

ISSN： 0166-3542

年卷期： 2008 年 79 卷 2 期

页码：

收录情况： SCI

摘要： Infectious bursal disease (IBD) leads to considerable economic losses for the poultry industry by inducing severe immunosuppression and high mortality in chickens. The objective of this study was to determine if RNA interference (RNAi) could be utilized to inhibit IBDV replication in vitro. We selected 3 short interfering RNA (siRNA) sequences (SiVp1(618), siVP1(1115), and SiVp1(2571)) based on conserved regions in the vp1 gene of the infectious bursal disease virus (IBDV). When the Vero cells were transfected with siRNA, synthesized via in vitro transcription, and then infected with IBDV, SiVp1(2571) was discovered to be the most effective site for inhibiting IBDV replication. For long-term expression of siRNA and due to its suitability for large-scale preparation, the mouse U6 promoter was amplified using primers designed according to the SiVp1(2571) sequence. The resulting products were then subcloned into pEGFP-C1 to construct the shRNA expression vector pEC2571-shRNA. The shRNA-transfected Vero cells were then infected with IBDV. As compared to the control, the inhibitory rate in the pEC2571-shRNA-transfected group was 87.4%. Indirect immunofluorescence and real-time polymerase chain reaction (PCR) confirmed that VP1 expression decreased at both the protein and RNA levels as compared to that in the controls. The results presented here indicate that DNA vector-based RNAi could effectively inhibit IBDV replication in vitro. (C) 2007 Elsevier B.V. All rights reserved.

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