Evaluation of the effects of small interfering RNAs on in vitro replication of feline herpesvirus-1.
OBJECTIVE
To evaluate the ability of 5 small interfering RNAs (siRNAs) targeting mRNA of the feline herpesvirus-1 (FHV-1) DNA polymerase gene to reduce in vitro viral replication and gene expression of FHV-1, to evaluate combinations of these siRNAs with siRNAs that target the glycoprotein D gene of FHV-1, and to determine the combination or combinations of siRNAs that yield the greatest inhibition of in vitro viral replication.


METHODS
Cultured Crandell-Rees feline kidney (CRFK) cells.


METHODS
CRFK cells were transfected with siRNAs designed to target mRNA of the FHV-1 DNA polymerase gene. Effective treatment was determined by quantification of the inhibition of mRNA available for DNA polymerase translation, viral protein production, and viral replication. Combinations of 2 siRNAs that target mRNA of the FHV-1 DNA polymerase gene and 2 siRNAs that target the mRNA of the essential FHV-1 glycoprotein D gene were evaluated for the ability to inhibit viral replication.


RESULTS
Verified by a reduction in viral gene expression, 2 of the 5 siRNAs designed to target mRNA of the FHV-1 DNA polymerase gene significantly suppressed viral replication. Two combinations of siRNAs that target mRNA of the FHV-1 DNA polymerase gene, the FHV-1 glycoprotein D gene, or both also significantly suppressed viral replication.


CONCLUSIONS
Combinations of siRNAs that target mRNA of the FHV-1 DNA polymerase gene, FHV-1 glycoprotein D gene, or both could potentially be used as a treatment for the prevention of clinical disease associated with FHV-1 infection.
Department of Comparative Medicine, College of Veterinary Medicine, University of Tennessee, Knoxville, TN 37996, USA.
Rebecca P
RP
Wilkes
Stephen A
SA
Kania
D000818:Animals; D015153:Blotting, Western / Q000662:veterinary; D002371:Cat Diseases / Q000821:virology*; D002415:Cats; D002460:Cell Line; D002470:Cell Survival / Q000235:genetics; D005434:Flow Cytometry / Q000662:veterinary; D015967:Gene Expression Regulation, Viral; D006564:Herpesviridae / Q000235:genetics / Q000502:physiology*; D006566:Herpesviridae Infections / Q000662:veterinary* / Q000821:virology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics*; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D014162:Transfection; D014759:Viral Envelope Proteins / Q000235:genetics; D014779:Virus Replication / Q000235:genetics*
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012367:RNA, Viral; D014759:Viral Envelope Proteins; C488986:glycoprotein D, Feline herpesvirus 1