April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Small RNAs Targeting ICP4 Affect HSV-1 Replication In Vitro
Author Affiliations & Notes
  • Kaili Wu
    Zhongshan Ophthalmic Center, State Key Laboratory of Ophthalmology, Sun Yat-Sen Univ, Guangzhou, China
  • Fang Duan
    Zhongshan Ophthalmic Center, State Key Laboratory of Ophthalmology, Sun Yat-Sen Univ, Guangzhou, China
  • Shiming Ni
    Zhongshan Ophthalmic Center, State Key Laboratory of Ophthalmology, Sun Yat-Sen Univ, Guangzhou, China
  • Yuhong Nie
    Zhongshan Ophthalmic Center, State Key Laboratory of Ophthalmology, Sun Yat-Sen Univ, Guangzhou, China
  • Qiang Huang
    Zhongshan Ophthalmic Center, State Key Laboratory of Ophthalmology, Sun Yat-Sen Univ, Guangzhou, China
  • Ziyan Chen
    Zhongshan Ophthalmic Center, State Key Laboratory of Ophthalmology, Sun Yat-Sen Univ, Guangzhou, China
  • Footnotes
    Commercial Relationships  Kaili Wu, None; Fang Duan, None; Shiming Ni, None; Yuhong Nie, None; Qiang Huang, None; Ziyan Chen, None
  • Footnotes
    Support  NSFC grant No.81070765
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 5803. doi:
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      Kaili Wu, Fang Duan, Shiming Ni, Yuhong Nie, Qiang Huang, Ziyan Chen; Small RNAs Targeting ICP4 Affect HSV-1 Replication In Vitro. Invest. Ophthalmol. Vis. Sci. 2011;52(14):5803.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : HSV-1 infected-cell polypeptide 4 (ICP4) gene plays key role during the virus infection. This study sought to investigate the effects of siRNAs and microRNA (miR-H6) targeting ICP4 on the lytic infection of HSV-1 in cultured cells.

Methods: : HSV-1 lytic infections were conducted in cultured human cornea epithelial cell(HCE), retinal pigment cell(RPE) and Vero cell. Small RNAs were transfected into cultured cells. The small RNAs’ effects were evaluated by western blot, plaque-reduction assays, indirect immunofluorescence and RT-PCR. The viral titre was detected by the TCID50 method.

Results: : Small RNA decreased ICP4 expression in cultured cells that were infected with HSV-1. Compared with HSV-1 infection alone or transfection with negative (control) small RNA, the viral titre and the cytopathic effect were significantly decreased in cultured cells transfected with ICP4-targeting small RNAs (siRNA and miR-H6). The siRNA effectively silenced HSV-1 ICP4 expression on both mRNA and the protein levels (p<0.05). miR-H6 also revealed the predominant inhibition on the HSV-1 production and the ICP4 expression in both HCE and RPE that were infected with the virus.

Conclusions: : ICP4-targeting siRNA and miR-H6 can inhibit HSV-1 replication in RPE and HCE cells. These provide a foundation for development of small RNA as an antiviral therapy.

Keywords: herpes simplex virus • cornea: epithelium • retinal pigment epithelium 
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