Actively replicating West Nile virus is resistant to cytoplasmic delivery of siRNA.
BACKGROUND
West Nile virus is an emerging human pathogen for which specific antiviral therapy has not been developed. Recent studies have suggested that RNA interference (RNAi) has therapeutic potential as a sequence specific inhibitor of viral infection. Here, we examine the ability of exogenous small interfering RNAs (siRNAs) to block the replication of West Nile virus in human cells.


RESULTS
WNV replication and infection was greatly reduced when siRNA were introduced by cytoplasmic-targeted transfection prior to but not after the establishment of viral replication. WNV appeared to evade rather than actively block the RNAi machinery, as sequence-specific reduction in protein expression of a heterologous transgene was still observed in WNV-infected cells. However, sequence-specific decreases in WNV RNA were observed in cells undergoing active viral replication when siRNA was transfected by an alternate method, electroporation.


CONCLUSIONS
Our results suggest that actively replicating WNV RNA may not be exposed to the cytoplasmic RNAi machinery. Thus, conventional lipid-based siRNA delivery systems may not be adequate for therapy against enveloped RNA viruses that replicate in specialized membrane compartments.
Department of Medicine, Washington University School of Medicine, 660 South Euclid Avenue, Box 8051, St Louis, MO 63110, USA. brian.geiss@colostate.edu
Brian J
BJ
Geiss
Theodore C
TC
Pierson
Michael S
MS
Diamond
D002460:Cell Line; D003593:Cytoplasm / Q000235:genetics / Q000821:virology; D018274:Electroporation; D006801:Humans; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics*; D011933:Reagent Kits, Diagnostic; D013997:Time Factors; D014162:Transfection; D014779:Virus Replication / Q000235:genetics; D014901:West Nile Fever / Q000628:therapy* / Q000821:virology; D014902:West Nile virus / Q000235:genetics / Q000502:physiology*
D034741:RNA, Small Interfering; D011933:Reagent Kits, Diagnostic