title: Artificial microRNAs can effectively inhibit replication of Venezuelan equine encephalitis virus.
abstract: Venezuelan equine encephalitis virus is a member of the alphavirus family and genus togaviridae. VEEV is highly infectious in aerosol form and has been weaponized in the past making it a potential biothreat agent. At present, there are no FDA approved antiviral treatments or vaccines for VEEV. Artificial microRNAs are small molecules which are expressed through endogenous microRNA machinery by RNA polymerase II. These artificial microRNAs effectively inhibit gene expression and are non-toxic to the host cell. VEEV RNA dependent RNA polymerase (RdRp) is central to VEEV replication. Therefore, we hypothesize that targeted inhibition of VEEV RdRp using artificial microRNAs may efficiently inhibit VEEV replication. Five artificial microRNAs were tested in vitro in BHK cells. Three of these artificial miRNAs showed significant inhibition of VEEV replication. Further, these microRNAs were cloned into the expression vector in combination to see the synergistic effect on VEEV replication. Combination of more than one miRNA did not result in significant inhibition of virus replication. In conclusion, we have shown that RNAi through artificial microRNAs effectively inhibits VEEV replication and is significantly less toxic in comparison to siRNAs.
authors.affiliation: Department of Pathology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USA; Department of Biological Sciences, Birla Institute of Technology and Science, Pilani, Rajasthan 333031, India.
authors.forename: Manish
authors.initials: M
authors.lastname: Bhomia
authors.forename: Anuj
authors.initials: A
authors.lastname: Sharma
authors.forename: Manoshi
authors.initials: M
authors.lastname: Gayen
authors.forename: Paridhi
authors.initials: P
authors.lastname: Gupta
authors.forename: Radha K
authors.initials: RK
authors.lastname: Maheshwari
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism*; D001688:Biological Products / Q000378:metabolism*; D002460:Cell Line; D006224:Cricetinae; D004666:Encephalitis Virus, Venezuelan Equine / Q000235:genetics / Q000502:physiology*; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism*; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000998:Antiviral Agents; D001688:Biological Products; D035683:MicroRNAs
keywords: Antiviral; Artificial microRNA; VEEV; nsp-4; siRNA