Authors: Chetan D Meshram Niraj K Singh Arvind A Sonwane Sachin S Pawar B P Mishra V K Chaturvedi Mohini Saini R P Singh Praveen K Gupta
Publish Date: 2013/06/11
Volume: 158, Issue: 11, Pages: 2323-2332
Abstract
Small interfering RNAs siRNAs targeting rabies virus RV glycoprotein G and nucleoprotein N genes were evaluated as antiviral agents against rabies virus in vitro in BHK21 cells To select effective siRNAs targeting RVG a plasmidbased transient cotransfection approach was used In this siRNAs were expressed as short hairpin RNAs shRNAs and their ability to inhibit RVG gene expression was evaluated in cells transfected with a plasmid expressing RVG The nine different siRNAs designed to target RVG exhibited varying degrees of knockdown of RVG gene expression One siRNA siG7 with considerable effect in knockdown of RVG expression also demonstrated significant inhibition of RV multiplication in BHK21 cells after in vitro challenge with the RV Pasteur virus11 PV11 strain A decrease in the number of fluorescent foci in siRNAtreated cells and a reduction 868 in the release of RV into infected cell culture supernatant indicated the antirabies potential of siRNA Similarly treatment with one siRNA targeting RVN resulted in a decrease in the number of fluorescent foci and a reduction 859 in the release of RV As a dual gene silencing approach where siRNAs targeting RVG and RVN genes were expressed from single construct the antirabiesvirus effect was observed as an 874 reduction in the release of RV These results demonstrate that siRNAs targeting RVG and N both in single and dual form have potential as antiviral agent against rabiesThis study was generously supported in part by the project “Evaluation of antirabies effect of small interfering RNA siRNA delivered through viral vector” Grant No BT/PR9941/AGR/ 36/22/2007 from Department of Biotechnology DBT Government of India and Indian Council of Agricultural Research ICAR India
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