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Inhibition of West Nile virus Replication by Bifunctional siRNA Targeting the NS2A and NS5 Conserved Region

[ Vol. 18 , Issue. 3 ]

Author(s):

Divyanshi Karothia, Paban Kumar Dash, Manmohan Parida, Sameer Bhagyawant and Jyoti S. Kumar*   Pages 180 - 190 ( 11 )

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.

Keywords:

siRNA, WNV, Bifunctional siRNA, Immunostimulatory siRNA, Flaviviruses, RT-PCR.

Affiliation:

Division of Virology, Defence Research and Development Establishment, Jhansi Road, Gwalior-474002, Division of Virology, Defence Research and Development Establishment, Jhansi Road, Gwalior-474002, Division of Food Microbiology, Defence Food Research Laboratory, Siddhartha Nagar, Mysore-570011, School of studies in Biotechnology, Jiwaji University, Gwalior, Division of Virology, Defence Research and Development Establishment, Jhansi Road, Gwalior-474002



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