ABSTRACT
OBJECTIVE: To construct a recombinant lentiviral vector containing integrin ß1 shRNA to provide an effective tool for integrin ß1 gene effect and a possible mechanism of Sombati cell of clinical refractory epilepsy. METHODS: Four lentiviral vectors containing integrin ß1 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 ß1 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. CONCLUSION: The recombinant lentiviral vector containing integrin ß1 shRNA is constructed successfully. And the gene-silencing effects are effective and stable in neonatal rat hippocampal neurons and Sombati cells.