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


METHODS
Crandell-Rees feline kidney cells.


METHODS
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
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.
Wilkes
Rebecca P
RP
Department of Comparative Medicine, College of Veterinary Medicine, University of Tennessee, Knoxville, TN 37996, USA.
Kania
Stephen A
SA
D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western / Q000662:veterinary; D002415:Cats; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D005434:Flow Cytometry / Q000662:veterinary; D005786:Gene Expression Regulation / Q000235:genetics*; D006386:Hemagglutination Tests / Q000662:veterinary; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D018140:Varicellovirus / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D010948:Viral Plaque Assay / Q000662:veterinary; D053585:Virus Attachment / Q000187:drug effects*
D017931:DNA Primers; D034741:RNA, Small Interfering; D014759:Viral Envelope Proteins; C488986:glycoprotein D, Feline herpesvirus 1