Short Interfering RNA-Mediated Inhibition of Herpes Simplex Virus Type 1 Gene Expression and Function during Infection of Human Keratinocytes

• DOI: 10.1128/jvi.78.19.10276-10281.2004
• Publication Date: 2004-09-14T20:48:09Z
• Authors: Prakash K. Bhuyan; Katalin Karikò; John Capodici; John Lubinski; Lauren M. Hook; Harvey M. Friedman; Drew Weissman
• Keywords: Gene Expression Regulation, Viral; Keratinocytes; 0301 basic medicine; Genes, Viral; Herpesvirus 1, Human; Viral Plaque Assay; Blotting, Northern; Flow Cytometry; Virus Replication; 3. Good health; 03 medical and health sciences; Viral Envelope Proteins; Humans; RNA, Viral; RNA Interference; RNA, Small Interfering; Cells, Cultured

ABSTRACTRNA 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.