title: Evaluation of single and dual siRNAs targeting rabies virus glycoprotein and nucleoprotein genes for inhibition of virus multiplication in vitro.
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 BHK-21 cells. To select effective siRNAs targeting RV-G, a plasmid-based transient co-transfection approach was used. In this, siRNAs were expressed as short hairpin RNAs (shRNAs), and their ability to inhibit RV-G gene expression was evaluated in cells transfected with a plasmid expressing RV-G. The nine different siRNAs designed to target RV-G exhibited varying degrees of knockdown of RV-G gene expression. One siRNA (si-G7) with considerable effect in knockdown of RV-G expression also demonstrated significant inhibition of RV multiplication in BHK-21 cells after in vitro challenge with the RV Pasteur virus-11 (PV-11) strain. A decrease in the number of fluorescent foci in siRNA-treated cells and a reduction (86.8 %) in the release of RV into infected cell culture supernatant indicated the anti-rabies potential of siRNA. Similarly, treatment with one siRNA targeting RV-N resulted in a decrease in the number of fluorescent foci and a reduction (85.9 %) in the release of RV. As a dual gene silencing approach where siRNAs targeting RV-G and RV-N genes were expressed from single construct, the anti-rabies-virus effect was observed as an 87.4 % reduction in the release of RV. These results demonstrate that siRNAs targeting RV-G and N, both in single and dual form, have potential as antiviral agent against rabies.
authors.affiliation: Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, 243 122, India.
authors.forename: Chetan D
authors.initials: CD
authors.lastname: Meshram
authors.forename: Niraj K
authors.initials: NK
authors.lastname: Singh
authors.forename: Arvind A
authors.initials: AA
authors.lastname: Sonwane
authors.forename: Sachin S
authors.initials: SS
authors.lastname: Pawar
authors.forename: B P
authors.initials: BP
authors.lastname: Mishra
authors.forename: V K
authors.initials: VK
authors.lastname: Chaturvedi
authors.forename: Mohini
authors.initials: M
authors.lastname: Saini
authors.forename: R P
authors.initials: RP
authors.lastname: Singh
authors.forename: Praveen K
authors.initials: PK
authors.lastname: Gupta
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000235:genetics* / Q000378:metabolism; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D006224:Cricetinae; D020868:Gene Silencing; D006023:Glycoproteins / Q000235:genetics* / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D019590:Nucleocapsid Proteins / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D011818:Rabies / Q000188:drug therapy / Q000821:virology; D011820:Rabies virus / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000956:Antigens, Viral; D000998:Antiviral Agents; D006023:Glycoproteins; D019590:Nucleocapsid Proteins; D034741:RNA, Small Interfering; D014759:Viral Envelope Proteins; C064951:glycoprotein G, Rabies virus; C099601:nucleocapsid protein, Rabies virus