Construction and identification of lentiviral vector of MMP-9 gene RNAi

Objective To construct lentiviral vector of matrix metalloproteinase 9(MMP-9) RNA interference(RNAi) of mice.Methods Towards the MMP-9 gene sequences of mice,in accordance with RNAi sequence design principles using the public Web site,RNAi target sequences were designed,then the target sequences of Oligo DNA were synthesized and annealed to double-stranded DNA,connecting with pGCL-GFP vector digested by Hpa Ⅰ and Xho Ⅰ.Short hairpin RNA lentiviral vectors were constructed.The positive clones identified by PCR and sequenced.The MMP-9 expression in the downward effect in 293T cells by Western blot was studied.Results The PCR identification and DNA sequencing showed that the lentiviral vector of MMP-9 shRNA-oligonucleotide chain was inserted correctly;and MMP-9 RNAi lentiviral vectors inhibited the expression of MMP-9.Conclusion A lentiviral vector of MMP-9 gene RNAi of mice is constructed successfully by the genetic engineering technology,and it provides a condition for gene therapy in vivo.