Construction and identification of ADAM17-targeting RNA interference lentivirus expressive vectors

Objective To construct recombinant lentiviral vectors for RNA interference(RNAi) targeting a disintegrin and metalloproteinase 17(ADAM17) gene and package recombinant lentivirus.Method Four pairs of short hairpin RNA(shRNA) sequences were designed according to the sequence of human ADAM17 gene.After synthesis and annealing,the double-stranded oligonucleotides were cloned into the pLVTHM vectors,which were confirmed using DNA sequencing analysis.The silencing effects of the ADAM17 siRNA expression vectors were analyzed.The most effective ADAM17 RNAi expression vector was packaged to recombinant lentivirus in 293T cells.The titer of lentivirus was detected by flow cytometry.Result DNA sequencing demonstrated that the recombinant lentiviral vectors were constructed successfully.The most effective ADAM17 siRNA expression vector was selected successfully.Then the recombinant lentivirus containing human ADAM17 gene was packaged successfully and its titer was 2.16×108 TU/mL.Conclusion The recombinant lentiviral vector targeting human ADAM17 gene was constructed,and the recombinant lentivirus was packaged successfully.