title: Vector-delivered artificial miRNA effectively inhibited replication of Chikungunya virus.
abstract: Chikungunya virus (CHIKV) has emerged as one of the most significant arboviral threats in many parts of the world. In spite of large scale morbidity, and long lasting polyarthralgia, no licensed vaccine or antivirals are available for the clinical management of CHIKV infection. In this study, a novel RNA interference based strategy has been adopted for effective inhibition of CHIKV. Four artificial microRNAs (amiRNAs) were designed to target different regions of CHIKV genome. These amiRNAs significantly inhibited CHIKV replication in Vero cells at both RNA and protein levels as assessed by qRT-PCR, immunoblotting and immunofluorescence techniques. Further inhibition of the infectious CHIKV up to 99.8% was demonstrated by plaque reduction assay. Concatemerization of amiRNA resulted in higher inhibition of CHIKV than individual amiRNAs. In addition, we studied the effect of combination of RNAi based therapy with other classical antivirals like chloroquine, ribavirin and mycophenolic acid, that helped in understanding the rational selection of RNAi based combination therapy. These findings provide a promising avenue for the development of novel amiRNA or combination based therapeutics against emerging CHIKV.
authors: Virology Division, Defence Research & Development Establishment, Gwalior, 474002, India.
authors: Amrita
authors: A
authors: Saha
authors: School of Studies in Biotechnology, Jiwaji University, Gwalior, India.
authors: Sameer S
authors: SS
authors: Bhagyawant
authors: Virology Division, Defence Research & Development Establishment, Gwalior, 474002, India.
authors: Manmohan
authors: M
authors: Parida
authors: Virology Division, Defence Research & Development Establishment, Gwalior, 474002, India. Electronic address: pabandash@drde.drdo.in.
authors: Paban Kumar
authors: PK
authors: Dash
mesh_terms: D000818:Animals; D000962:Antimalarials / Q000494:pharmacology; D000998:Antiviral Agents / Q000494:pharmacology; D002646:Chikungunya virus / Q000187:drug effects / Q000502:physiology; D002522:Chlorocebus aethiops; D002738:Chloroquine / Q000494:pharmacology; D004261:DNA Replication / Q000187:drug effects; D005822:Genetic Vectors; D035683:MicroRNAs / Q000138:chemical synthesis* / Q000235:genetics*; D034622:RNA Interference; D012254:Ribavirin / Q000494:pharmacology; D014162:Transfection; D014709:Vero Cells; D010948:Viral Plaque Assay; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000962:Antimalarials; D000998:Antiviral Agents; D035683:MicroRNAs; D012254:Ribavirin; D002738:Chloroquine
keywords: Antiviral drugs; Artificial miRNA; Chikungunya virus; GFP; RNAi