Short interfering RNA-mediated inhibition of herpes simplex virus type 1 gene expression and function during infection of human keratinocytes.
RNA interference (RNAi) is an antiviral mechanism that is activated when double-stranded RNA is cleaved into fragments, called short interfering RNA (siRNA), that prime an inducible gene silencing enzyme complex. We applied RNAi against a herpes simplex virus type 1 (HSV-1) gene, glycoprotein E, which mediates cell-to-cell spread and immune evasion. In an in vitro model of infection, human keratinocytes were transfected with siRNA specific for glycoprotein E and then infected with wild-type HSV-1. RNAi-mediated gene silencing reproduced the small plaque phenotype of a gE-deletion mutant virus. The specificity of gene targeting was demonstrated by flow cytometry and Northern blot analyses. Exogenous siRNA can suppress HSV-1 glycoprotein E expression and function during active infection in vitro through RNAi. This work establishes RNAi as a genetic tool for the study of HSV and provides a foundation for development of RNAi as a novel antiviral therapy.
Bhuyan
Prakash K
PK
Division of Infectious Diseases, Department of Medicine, University of Pennsylvania, Philadelphia, PA, USA. bhuyan@prodigy.net
Karikò
Katalin
K
Capodici
John
J
Lubinski
John
J
Hook
Lauren M
LM
Friedman
Harvey M
HM
Weissman
Drew
D
D015152:Blotting, Northern; D002478:Cells, Cultured; D005434:Flow Cytometry; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral; D018259:Herpesvirus 1, Human / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D015603:Keratinocytes / Q000821:virology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000502:physiology*; D012367:RNA, Viral / Q000032:analysis; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D010948:Viral Plaque Assay; D014779:Virus Replication / Q000187:drug effects
D034741:RNA, Small Interfering; D012367:RNA, Viral; D014759:Viral Envelope Proteins; C044768:glycoprotein E, herpes simplex virus type 1