title: [Construction of recombinant lentiviral vector containing integrin β1 shRNA and its effects on integrin β1 gene expression in Sombati cell culture model of refractory epilepsy].
abstract: OBJECTIVE
  To construct a recombinant lentiviral vector containing integrin β1 shRNA to provide an effective tool for integrin β1 gene effect and a possible mechanism of Sombati cell of clinical refractory epilepsy.
  
  
  METHODS
  Four lentiviral vectors containing integrin β1 shRNA were constructed and transfected into 293T cells. PC12 cells were infected by concentrated lentivirus and the gene-silencing efficiency was verified. And the most effective lentivirus containing shRNA was selected with Western blot. Then neonatal rat hippocampal neurons and Sombati cells were infected by lentivirus containing shRNA and the gene-silencing efficiency was also monitored by Western blot.
  
  
  RESULTS
  RNAi lentivirus expression vectors targeting rat integrin β1 gene were successfully constructed and confirmed by DNA sequencing. The recombinant lentivirus particles were packaged successfully to produce a sufficient titer for subsequent experiments. The expression of protein significantly decreased in rat hippocampal neurons and rat Sombati cells after vector transfection.
  
  
  CONCLUSIONS
  The recombinant lentiviral vector containing integrin β1 shRNA is constructed successfully. And the gene-silencing effects are effective and stable in neonatal rat hippocampal neurons and Sombati cells.
authors.affiliation: Department of Neurology, First Affiliated Hospital, Guangxi Medical University, Nanning 530021, China.
authors.forename: Jie-mei
authors.initials: JM
authors.lastname: Ye
authors.forename: Yuan
authors.initials: Y
authors.lastname: Wu
authors.forename: Jin-shan
authors.initials: JS
authors.lastname: Huang
authors.forename: Si-jun
authors.initials: SJ
authors.lastname: Li
authors.forename: Yun
authors.initials: Y
authors.lastname: Liu
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D018929:Cell Culture Techniques; D004827:Epilepsy / Q000628:therapy*; D015870:Gene Expression; D005822:Genetic Vectors; D019012:Integrin beta1 / Q000235:genetics*; D016086:Lentivirus / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000627:therapeutic use*; D051381:Rats; D014162:Transfection
chemical_list: D019012:Integrin beta1; D034741:RNA, Small Interfering