title: Short hairpin RNA-mediated inhibition of HSV-1 gene expression and function during HSV-1 infection in Vero cells.
abstract: OBJECTIVE
  To evaluate the efficiency of 3 short hairpin RNA (shRNA) interfering with the herpes simplex virus type 1 (HSV-1) gene coding glycoprotein D (gD) for inhibiting the gD expression and virus replication in vitro.
  
  
  METHODS
  Vero cells were selected for an in vitro model of infection. Three shRNA sequences (shRNAgD1, -gD2, and -gD3) targeting specifically the gD gene of HSV-1 were selected for evaluating the antiviral effects. The antiviral effects of shRNA in the cells infected with HSV-1 were evaluated by cytopathic effect (CPE) observations and plaque assays. The transcription level of viral RNA and the gD expression were studied by RT-PCR, Western blotting, and flow cytometry.
  
  
  RESULTS
  With the 3 shRNA at a final concentration of 120 nmol/L, a significant inhibition of CPE in the HSV-1-infected cells was observed. The ED50 of shRNA-gD1, gD2, and gD3 were 48.74+/-2.57, 57.13+/-3.24, and 114.64+/-5.12 nmol/L, respectively. The gD gene decreased significantly after viral infection in the Vero cells pretreated with shRNA compared to the virus group. The expressions of the gD protein, determined by Western blotting and flow cytometry, were also drastically decreased in shRNA-transfected cells.
  
  
  CONCLUSIONS
  Exogenous shRNA molecules can suppress the HSV-1 gD expression. They are inhibitors of HSV replication during infection in Vero cells.
authors.affiliation: State Key Laboratory of Virology, Institute of Medical Virology, School of Medicine, Wuhan University, Wuhan 430071, China.
authors.forename: Yuan-yuan
authors.initials: YY
authors.lastname: Liu
authors.forename: Hai-ying
authors.initials: HY
authors.lastname: Deng
authors.forename: Guang
authors.initials: G
authors.lastname: Yang
authors.forename: Wen-lin
authors.initials: WL
authors.lastname: Jiang
authors.forename: Laurent
authors.initials: L
authors.lastname: Grossin
authors.forename: Zhan-qiu
authors.initials: ZQ
authors.lastname: Yang
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D003588:Cytopathogenic Effect, Viral / Q000187:drug effects; D015967:Gene Expression Regulation, Viral / Q000235:genetics / Q000502:physiology*; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000502:physiology*; D055029:Inverted Repeat Sequences / Q000235:genetics; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics; D014779:Virus Replication / Q000187:drug effects
chemical_list: D012367:RNA, Viral; D014759:Viral Envelope Proteins; C039447:glycoprotein D, Human herpesvirus 1