摘要
背景:西尼罗河病毒(WNV)已成为世界上许多地区最重要的虫媒病毒感染之一,并且与脑炎主要影响人类和马匹有关。尽管WNV对公众健康构成威胁,但是没有可用于治疗WNV的疫苗或治疗剂。 方法:在这项研究中,我们测试了一种新的基于RNA干扰的技术来抑制Vero细胞中的WNV复制。针对WNV的NS2A和NS5区设计了两种siRNA,其在黄病毒中高度保守,因为它分别在细胞凋亡和病毒复制中起重要作用。除此之外,通过将免疫刺激基序与NS2A和NS5特异性siRNA连接来设计双功能siRNA。通过检测感染性病毒及其基因组来评估抗病毒活性。 结果:与对照相比,双功能siRNA导致siRNA转染细胞中病毒滴度的显着降低。抗病毒效力在感染后48小时最有效。这些结果与定量RT-PCR测定一致,揭示了WNV基因组RNA的类似减少。管家基因的表达不受siRNA的影响,表明没有脱靶效应和不干扰细胞机制。 结论:因此,这种双功能siRNA干预为治疗WNV疾病铺平了道路
关键词: siRNA,WNV,双功能siRNA,免疫刺激性siRNA,黄病毒,RT-PCR。
Current Gene Therapy
Title:Inhibition of West Nile virus Replication by Bifunctional siRNA Targeting the NS2A and NS5 Conserved Region
Volume: 18 Issue: 3
关键词: siRNA,WNV,双功能siRNA,免疫刺激性siRNA,黄病毒,RT-PCR。
摘要: Background: The West Nile Virus (WNV) has emerged as one of the most significant arboviral infection in many parts of the world and is associated with the encephalitis affecting mainly human and horses. In spite of the fact that the WNV is threat for the public health, there is no vaccine or therapeutic available for the treatment of WNV.
Methods: In this study, we tested a novel RNA interference based technique to inhibit WNV replication in Vero cells. Two siRNAs were designed against the NS2A and NS5 regions of WNV which are highly conserved among Flaviviruses as it play important role in apoptosis and in viral replication respectively. In addition to this, dual functional siRNA is designed by joining an immunostimulatroy motif with the NS2A and NS5 specific siRNA. The antiviral activity was evaluated by detecting both the infectious virus and its genome.
Results: The bifunctional siRNA resulted in significant reduction of virus titre in siRNA transfected cells as compared to controls. The antiviral efficacy was most effective at 48hr post infection. These results were in accordance with the quantitative RT-PCR assay revealing similar reduction in WNV genomic RNA. The expression of housekeeping gene was not affected by the siRNA indicating no off target effect and non-interference in cellular mechanism.
Conclusion: Thus, this bifunctional siRNA intervention paves the new way for therapeutic treatment of WNV disease.
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Cite this article as:
Inhibition of West Nile virus Replication by Bifunctional siRNA Targeting the NS2A and NS5 Conserved Region, Current Gene Therapy 2018; 18 (3) . https://dx.doi.org/10.2174/1566523218666180607091311
DOI https://dx.doi.org/10.2174/1566523218666180607091311 |
Print ISSN 1566-5232 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5631 |
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