Effect of lentivirus-mediated RNA interference on expression of / 中华麻醉学杂志

ABSTRACT

Objective To investigate the effect of lentivirus-mediated RNA interference(RNAi) on the expression of PKCγ mRNA and protein in rat neurons.Methods Primary cultured cortical neurons from SD rat (16 days of pregnency) embryos were randomly divided into 3 groups with 6 wells in each groupcontrol group (group C),negative group(group NC)and RNAi group.Group C received no treatment.Each well in group NC was given negative lentivirus 3 × 105.Each well in group RNAi was given the recombinant lentiviral vector containing PKCγ shRNA(LV-PKC7 shRNA).The expression of PKCT mRNA and protein in rat neurons was detected by RT-PCR and Western blot 5 days later.The interference efficiency was ealculated.Results Compared with group NC,the expression of PKCγ mRNA and protein was down-regulatedin group RNAi(P<0.05),but no significant change Was found in group NC(P>0.05).The interference efficiency of gene and protein were 99.3%and 85.2%respectively.Conclusion Lentivirus-mediated RNAi can down restate the expression of PKC7 gene and protein in rat neurons.