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Reduction of infectious bursal disease virus replication by shRNAs targeting the VP1 and VP2 genes driven by chicken U6 promoter

文献类型： 外文期刊

作者： Wei Ouyang 1 ; Ma, Jin-rong 1 ; Wang, Yong-qiang 2 ; Qin, Li-ting 2 ; Jiang, Jie-yuan 1 ; Fan, Hong-jie 3 ; Wang, Xiao 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Vet Med, Minist Agr, Key Lab Vet Biol Engn & Technol, Nanjing 210014, Jiangsu, Peoples R China

2.Chinese Acad Agr Sci, Harbin Vet Res Inst, State Key Lab Vet Biotechnol, Harbin 150001, Peoples R China

3.Nanjing Agr Univ, Coll Vet Med, Dept Microbiol & Immunol, Harbin 210095, Peoples R China

关键词： Infectious bursal disease virus;RNA interference;Short-hairpin RNA;Chicken U6 promoter

期刊名称：VETERINARY MICROBIOLOGY （ 影响因子：3.293； 五年影响因子：3.599 ）

ISSN： 0378-1135

年卷期： 2013 年 162 卷 1 期

页码：

收录情况： SCI

摘要： Infectious bursal disease virus (IBDV) causes a highly contagious and immunosuppressive disease in young chickens and results in considerable economic losses for the poultry industry. To suppress the replication of IBDV, two short hairpin RNAs (shRNAs) were designed for targeting the VP1 and VP2 genes of IBDV. Recombinant plasmids carrying each shRNA or two shRNAs were constructed based on vector pSilencer2.1-U6 in which the human U6 promoter was replaced with chicken U6 promoter. In chicken embryo fibroblasts, transfection with these shRNA plasmids 24 h before infection with IBDV B87 reduced 50% tissue culture infectious doses (TCID50) from 10(8.75) TCID50/0.1 mL to 10(3.75)-10(1.0) TCID50/0.1 mL. In 10-day old specific pathogen-free (SPF) chicken embryos, incubation with a mixture of IBDV B87 and a shRNA plasmid via the allantoic cavity resulted in 100% mortality and high IBDV virus titer in the control group but 25-0% mortality and near normal embryo development in the specific shRNA groups; additionally, IBDV VP1 and VP2 mRNA levels were reduced by 72-95% in the shRNA groups as compared with the control groups. When challenged with a virulent strain IBDV GX8/99, 14-day-old chickens pre-treated with the single shRNA plasmids or the dual shRNA plasmid showed approximately 70% or 90% survival at 5 days post-challenge while those pre-treated with control plasmid or saline had less than 5% survival. The current study suggests that two IBDV shRNAs expressed by a plasmid under chicken U6 promoter could effectively and synergistically reduce IBDV replication in vitro and in vivo. (C) 2012 Elsevier B.V. All rights reserved:

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