title: Inhibition of UL54 and UL97 genes of human cytomegalovirus by RNA interference.
abstract: Short interfering RNAs (siRNAs), namely siUL54-1 and siU54-2 targeting UL54 (DNA polymerase) gene, and siUL97-1 and siUL97-2 targeting UL97 (phosphotransferase) gene, were used to inhibit respective genes of Human cytomegalovirus (HCMV) and consequently the virus infection process in human foreskin fibroblast (HFF) cultures. The virus infection was monitored by cell morphology (CPE), levels of UL83 and IE86 mRNAs, and virus antigen. The results showed that siUL97-2 remarkably inhibited viral CPE while other siRNAs were less inhibitory. The siRNAs reduced the levels of UL83 mRNA but not that of IE86 mRNA; again, siUL97-2 was most inhibitory. Particularly, siUL97-2 reduced the UL83 mRNA level 14, 19, 203, and 37 times at 24, 48, 72, and 96 hrs post infection (p.i.), respectively. When tested for the effect on viral antigen by immunofluorescent assay (IFA), UL97-2 exerted a marked inhibition. These results demonstrate the effectiveness of siRNAs against experimental HCMV infection and indicate their therapeutic potential.
authors.affiliation: Department of Microbiology, College of Medicine, Catholic Hemopoietic Stem Cell Transplantation Center, The Catholic University of Korea, Seoul 137-701, South Korea.
authors.forename: M-C
authors.initials: MC
authors.lastname: Shin
authors.forename: S-K
authors.initials: SK
authors.lastname: Hong
authors.forename: J-S
authors.initials: JS
authors.lastname: Yoon
authors.forename: S-S
authors.initials: SS
authors.lastname: Park
authors.forename: S-G
authors.initials: SG
authors.lastname: Lee
authors.forename: D-G
authors.initials: DG
authors.lastname: Lee
authors.forename: W-S
authors.initials: WS
authors.lastname: Min
authors.forename: W-S
authors.initials: WS
authors.lastname: Shin
authors.forename: S-Y
authors.initials: SY
authors.lastname: Paik
mesh_terms: D000956:Antigens, Viral / Q000032:analysis; D000998:Antiviral Agents / Q000494:pharmacology; D002460:Cell Line; D003587:Cytomegalovirus / Q000187:drug effects / Q000235:genetics* / Q000502:physiology*; D003588:Cytopathogenic Effect, Viral; D004259:DNA-Directed DNA Polymerase / Q000096:biosynthesis / Q000235:genetics*; D005347:Fibroblasts / Q000821:virology; D019085:Fluorescent Antibody Technique, Direct; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D006801:Humans; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000096:biosynthesis / Q000235:genetics*; D034622:RNA Interference; D012333:RNA, Messenger / Q000032:analysis; D034741:RNA, Small Interfering / Q000494:pharmacology*; D012367:RNA, Viral / Q000032:analysis; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics*; D014779:Virus Replication / Q000187:drug effects / Q000235:genetics
chemical_list: D000956:Antigens, Viral; D000998:Antiviral Agents; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012367:RNA, Viral; C109367:UL54 protein, Human herpesvirus 5; D014764:Viral Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); C075365:ganciclovir kinase; D004259:DNA-Directed DNA Polymerase