Divyanshi Karothia 1 , Paban Kumar Dash 1 , Manmohan Parida 2 , Sameer Bhagyawant 3 , Jyoti S Kumar 1 Show Affiliations »
Abstract
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. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.
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 . Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.
Entities: Disease
Species
Keywords:
Bifunctional siRNA; Flaviviruses; Immunostimulatory siRNA; RT-PCR; WNV; siRNA.
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Year: 2018
PMID： 29874999 DOI： 10.2174/1566523218666180607091311
Source DB: PubMed Journal: Curr Gene Ther ISSN： 1566-5232 Impact factor: 4.391