title: Specific interference shRNA-expressing plasmids inhibit Hantaan virus infection in vitro and in vivo.
abstract: OBJECTIVE
  To investigate the antiviral effects of vectors expressing specific short hairpin RNAs (shRNAs) against Hantaan virus (HTNV) infection in vitro and in vivo.
  
  
  METHODS
  Based on the effects of 4 shRNAs targeting different regions of HTNV genomic RNA on viral replication, the most effective RNA interference fragments of the S and M genes were constructed in pSilencer-3.0-H1 vectors, and designated pSilencer-S and pSilencer-M, respectively. The antiviral effect of pSilencer-S/M against HTNV was evaluated in both HTNV-infected Vero-E6 cells and mice.
  
  
  RESULTS
  In HTNV-infected Vero-E6 cells, pSilencer-S and pSilencer-M targeted the viral nucleocapsid proteins and envelope glycoproteins, respectively, as revealed in the immunofluorescence assay. Transfection with pSilencer-S or pSilencer-M (1, 2, 4 μg) markedly inhibited the viral antigen expression in dose- and time-dependent manners. Transfection with either plasmid (2 μg) significantly decreased HTNV-RNA level at 3 day postinfectin (dpi) and the progeny virus titer at 5 dpi. In mice infected with lethal doses of HTNV, intraperitoneal injection of pSilencer-S or pSilencer-M (30 μg) considerably increased the survival rates and mean time to death, and significantly reduced the mean virus yields and viral RNA level, and alleviated virus-induced pathological lesions in lungs, brains and kidneys.
  
  
  CONCLUSIONS
  Plasmid-based shRNAs potently inhibit HTNV replication in vitro and in vivo. Our results provide a basis for development of shRNA as therapeutics for HTNV infections in humans.
authors.affiliation: State Key Laboratory of Virology/Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Yuan-yuan
authors.initials: YY
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Virology/Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Liang-jun
authors.initials: LJ
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Virology/Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhong
authors.affiliation: State Key Laboratory of Virology/Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Meng-xin
authors.initials: MX
authors.lastname: Shen
authors.affiliation: State Key Laboratory of Virology/Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Nian
authors.initials: N
authors.lastname: Ma
authors.affiliation: State Key Laboratory of Virology/Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Bing-yu
authors.initials: BY
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Virology/Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Luo
authors.affiliation: State Key Laboratory of Virology/Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Hou
authors.affiliation: State Key Laboratory of Virology/Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Zhan-qiu
authors.initials: ZQ
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Virology/Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Hai-rong
authors.initials: HR
authors.lastname: Xiong
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D018063:Hantaan virus / Q000502:physiology*; D006480:Hemorrhagic Fever with Renal Syndrome / Q000235:genetics / Q000628:therapy* / Q000821:virology; D008807:Mice, Inbred BALB C; D010957:Plasmids; D034741:RNA, Small Interfering / Q000235:genetics*; D014709:Vero Cells; D014779:Virus Replication
chemical_list: D034741:RNA, Small Interfering