title: Recombinant cell lines expressing shRNA targeting herpes simplex virus 2 VP16 inhibit virus replication.
abstract: OBJECTIVE
  To establish HSV2 VP16 targeting shRNA-expressing cell lines and investigate the antiviral effect of shRNA targeting HSV2 VP16.
  
  
  METHODS
  The cell lines Vero-shRNAs and negative-control Vero-shCON were established. Their inhibition effects on VP16 mRNA expression were tested by real-time fluorescent quantitative polymerase chain reaction (PCR) and their antiviral effects were evaluated by yield reduction assay. The influence of passage numbers on the inhibition ability of cell lines was researched.
  
  
  RESULTS
  Vero-shRNA24 targeting the upper stream, Vero-shRNA642 targeting the lower stream and Vero-shCON were established. Vero-shRNA24, Vero-shRNA642 and Vero-shRNA24 + 642 could reduce the VP16 mRNA significantly. Vero-shRNA24 was the most efficient. The HSV2 titers in Vero and Vero-shCON were the highest at 72 h after infection, and started decreasing thereafter. The viral titers of the Vero-shRNA groups reached a peak after 84 h and the highest titers were lower than in the Vero group. The inhibiting effect on VP16 mRNA expression and viral replication of Vero-shRNA24 cell lines of passages 10 and 20 were not significantly different from the primary cell line. Although of no statistical significance, the passage 50 cell line showed decreased inhibiting ability.
  
  
  CONCLUSIONS
  Recombinant cell lines expressing shRNA targeting HSV2 VP16 were established. They can stably inhibit HSV2 VP16 mRNA expression and viral replication within passage 50.
authors.affiliation: Department of Neurology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, PR China.
authors.forename: Rui
authors.initials: R
authors.lastname: Zhang
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.forename: Bo
authors.initials: B
authors.lastname: Song
authors.forename: Zhi Qiang
authors.initials: ZQ
authors.lastname: Han
authors.forename: Yu Ming
authors.initials: YM
authors.lastname: Xu
mesh_terms: D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D017405:Herpes Simplex Virus Protein Vmw65 / Q000235:genetics*; D018258:Herpesvirus 2, Human / Q000235:genetics* / Q000502:physiology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000096:biosynthesis / Q000235:genetics*; D014709:Vero Cells; D014779:Virus Replication / Q000235:genetics*
chemical_list: D017405:Herpes Simplex Virus Protein Vmw65; D034741:RNA, Small Interfering