title: [Establishment of a human embryonic stem cell line with stable β-catenin silencing by shRNA lentivirus].
abstract: OBJECTIVE
  To establish a human embryonic stem cell line with stably β-catenin gene silencing by lentivirus-mediated shRNA interference.
  
  
  METHODS
  PLKO.1-puro-β-catenin vector, a lentivirus plasmid expressing β-catenin shRNA, was packaged into 293FT cells. Human embryonic stem cells were infected with the lentivirus and the cell clones stably expressing β-catenin shRNA were selected by puromycin, with the uninfected cells and cells infected with the empty vector as the control. Real-time RT-PCR was used to evaluate the efficiency of β-catenin knocked down; β-catenin and OCT4 protein expression in the infected cells was examined using immunofluorescence assay.
  
  
  RESULTS
  Infection with β-catenin-specific shRNA lentivirus resulted in stable interference of β-catenin expression in human embryonic stem cells, which showed a β-catenin mRNA expression of only 1% of that in the uninfected cells. Infection with the empty vector produced no obvious effect on β-catenin expression compared to the uninfected cells. In the cells infected with β-catenin shRNA lentivirus, β-catenin protein expression was almost undetectable in immunofluorescence assay, while OCT4 was still expressed after the interference.
  
  
  CONCLUSIONS
  Lentiviral vector-delivered shRNA results in effective and stable β-catenin gene silencing in human embryonic stem cells.
authors.affiliation: Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha, China. applesy@gmail.com
authors.forename: Yi
authors.initials: Y
authors.lastname: Sun
authors.forename: Sicong
authors.initials: S
authors.lastname: Zeng
authors.forename: Guangxiu
authors.initials: G
authors.lastname: Lu
authors.forename: Ge
authors.initials: G
authors.lastname: Lin
mesh_terms: D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D010957:Plasmids; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection; D060449:Wnt Signaling Pathway; D051176:beta Catenin / Q000235:genetics* / Q000378:metabolism
chemical_list: C495270:CTNNB1 protein, human; D034741:RNA, Small Interfering; D051176:beta Catenin