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Silencing the respiratory syncytial virus fusion protein gene using RNA interference
Author(s) -
Vig Komal,
Lewis Nuruddeen,
Moore Eddie G.,
Pillai Sheekumar,
Dennis Vida A.,
Singh Shree Ram
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.603.7
Subject(s) - gene silencing , small interfering rna , rna interference , biology , virus , gene expression , virology , rna silencing , microbiology and biotechnology , gene , transfection , rna , genetics
Respiratory Syncytial Virus (RSV) is an important respiratory pathogen of medical significance that causes high mortality in infants. At present, there is no vaccine or antiviral drug against RSV. The fusion (F) protein of RSV is a good target for therapeutic purposes as it is primarily responsible for penetration of the virus into host cells and for subsequent syncytium formation during infection. In the present study, four small interfering RNAs (siRNAs), targeting different regions of the RSV F protein were used to silence the RSV F gene in order to disable membrane fusion, viral penetration and syncytium formation. Silencing of F gene was validated by using indirect immunofluorescence, western blot and RT‐PCR. We also analyzed the relationship between siRNA design and target structure. Our results show by cytoplasmic fluorescence and at 20 nM siRNA concentrations the effective silencing of the RSV F gene by the smartpool siRNA and the individual siRNA. RT‐PCR analysis also revealed a decrease in the F mRNA expression. Among the four siRNAs used, siRNA‐1 was most efficient resulting in complete silencing of the F gene expression at 5 nM siRNA concentration. The RSV mRNA target region had substantial effect on siRNA efficiency. The present study shows the silencing of F gene using siRNA and its possible therapeutic use. Financial support for this work by National Institutes of Health Grants 2S06 GM008219‐200012 and NSF‐CREST (HRD# 0734232 ) is acknowledged.