title: Protection of mice against lethal rabies virus challenge using short interfering RNAs (siRNAs) delivered through lentiviral vector.
abstract: The antiviral potential of small interfering RNAs (siRNAs) targeting rabies virus (RV) polymerase (L) and nucleoprotein (N) genes delivered through lentiviral vector was investigated. For in vitro evaluation, siRNAs expressing BHK-21 cell lines (BHK-L and BHK-N) were developed using transduction with Lenti-L and Lenti-N lentiviruses encoding siRNAs against RV-L and N genes, respectively. When these cell lines were challenged in vitro with RV Pasteur virus-11 (PV-11) strain, there was reduction in number of RV-specific foci and target gene transcripts indicating inhibitory effect on RV multiplication. For in vivo evaluation, mice were treated intracerebrally with lentiviruses and challenged with 20 LD50 of RV challenge virus standard-11 (CVS-11) strain by intramuscular route in masseter muscle. Five out of eight mice treated with Lenti-N survived indicating 62.5 % protection. The control and Lenti-L-treated mice died within 7-10 days indicating lethal nature of challenge virus and no protection. These results demonstrated that siRNA targeting RV-N could not only inhibit RV multiplication, but also conferred protection in mice against lethal RV challenge. These findings have implication on therapeutic use of siRNA targeting RV-N against RV infection.
authors.affiliation: Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, 243122, India.
authors.forename: Niraj K
authors.initials: NK
authors.lastname: Singh
authors.forename: Chetan D
authors.initials: CD
authors.lastname: Meshram
authors.forename: Arvind A
authors.initials: AA
authors.lastname: Sonwane
authors.forename: Shyam S
authors.initials: SS
authors.lastname: Dahiya
authors.forename: Sachin S
authors.initials: SS
authors.lastname: Pawar
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; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D006224:Cricetinae; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors / Q000008:administration & dosage; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000494:pharmacology*; D011818:Rabies / Q000628:therapy; D011820:Rabies virus / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D019562:Viral Load; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D019590:Nucleocapsid Proteins; D034741:RNA, Small Interfering; D014764:Viral Proteins; C097625:L protein, Rabies virus; D012321:DNA-Directed RNA Polymerases