Use of interfering RNAs targeted against feline herpesvirus 1 glycoprotein D for inhibition of feline herpesvirus 1 infection of feline kidney cells.

Abstract

OBJECTIVE:

To evaluate the use of RNA interference targeted against feline herpesvirus 1 (FHV-1) glycoprotein D for inhibition of FHV-1 infection of feline kidney cells.

SAMPLE POPULATION:

Crandell-Rees feline kidney cells.

PROCEDURES:

Crandell-Rees feline kidney cells were transfected with small interfering RNAs (siRNAs) that were designed to inhibit expression of FHV-1 glycoprotein D. The effectiveness of the treatment was determined via measurement of amounts of glycoprotein D mRNA, intracellular glycoprotein D, and glycoprotein D expressed on the surface of infected cells and comparison with appropriate control sample data.

RESULTS:

2 of 6 siRNAs tested were highly effective in reducing expression (ie, knockdown) of glycoprotein D mRNA; there were 77% and 85% reductions in mRNA in treated samples, compared with findings in the control samples. The knockdown of glycoprotein D mRNA resulted in reduced glycoprotein D protein production, as evidenced by 27% and 43% decreases in expression of glycoprotein D on the surface of siRNA-treated, FHV-1-infected cells and decreased expression of the protein within infected cells, compared with control samples. Treatment with these siRNAs also resulted in inhibition of FHV-1 replication, with reductions of 84% and 77% in amounts of virus released into cell culture supernatant, compared with findings in control samples.

CONCLUSIONS AND CLINICAL RELEVANCE:

2 chemically produced siRNAs that targeted the glycoprotein D gene significantly reduced FHV-1 titers in treated cells, suggesting that glycoprotein D is necessary for production of infective virions. This gene is a potential target for RNA interference as a means of inhibition of FHV-1 infection of feline cells.

• Authors: Rebecca P Wilkes, Stephen A Kania
• Journal: American journal of veterinary research (Am J Vet Res) Vol. 70 Issue 8 Pg. 1018-25 (Aug 2009) ISSN: 0002-9645 [Print] United States
• PMID: 19645584 (Publication Type: Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• DNA Primers
• RNA, Small Interfering
• Viral Envelope Proteins
• glycoprotein D, Feline herpesvirus 1

Topics

• Analysis of Variance
• Animals
• Blotting, Western (veterinary)
• Cats
• Cell Line
• DNA Primers (genetics)
• Flow Cytometry (veterinary)
• Gene Expression Regulation (genetics)
• Hemagglutination Tests (veterinary)
• RNA Interference
• RNA, Small Interfering (genetics, pharmacology)
• Reverse Transcriptase Polymerase Chain Reaction (veterinary)
• Varicellovirus (genetics)
• Viral Envelope Proteins (antagonists & inhibitors, genetics)
• Viral Plaque Assay (veterinary)
• Virus Attachment (drug effects)