title: [Construction of plasmid stably expressing shRNA and inhibition of HBV replication and expression].
abstract: OBJECTIVE
  To construct the vector stably expressing small hair RNA(shRNA) and investigate the effect of shRNA expressed with DNA vector on HBV replication and expression.
  
  
  METHODS
  The vector expressing shRNA designated pU6 was constructed by U6 promoter. The sequence of shRNA targeting on specific region of HBV genome against HBV was designed and synthesized, then cloned into pU6. The plasmid designated pU6B/HBVi was transfected into 2.2.15 cells using Lipofectamine 2000 reagent. The HBsAg and HBeAg in the supernatants of the transfected cells were measured by ELISA. The HBV copies in the transfected cells were measured by quantitative PCR.
  
  
  RESULTS
  Compared with controls, not only concentration of HBsAg and HBeAg but also HBV copies in transfected 2.2.15 cells decreased significantly by the plasmid expressing shRNA against HBV.
  
  
  CONCLUSIONS
  HBV replication and expression in 2.2.15 cells are inhibited by stably expressed shRNA.
authors.affiliation: Department of Medical Research, Guangzhou Army General Hospital, Guangzhou, China. zhangwater@tom.com
authors.forename: Hong-Bin
authors.initials: HB
authors.lastname: Zhang
authors.forename: Jie
authors.initials: J
authors.lastname: Wu
authors.forename: Wen-Ling
authors.initials: WL
authors.lastname: Zheng
authors.forename: Wen-Li
authors.initials: WL
authors.lastname: Ma
authors.forename: Jiang
authors.initials: J
authors.lastname: Xian
authors.forename: Wen-Yue
authors.initials: WY
authors.lastname: Xie
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: Jie
authors.initials: J
authors.lastname: Wang
mesh_terms: D000818:Animals; D002460:Cell Line; D017077:Culture Media, Conditioned / Q000378:metabolism; D015967:Gene Expression Regulation, Viral / Q000235:genetics*; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D055029:Inverted Repeat Sequences; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics*
chemical_list: D017077:Culture Media, Conditioned; D006514:Hepatitis B Surface Antigens; D034741:RNA, Small Interfering