Construction and package of recombinant lentivirus expressing shRNA targeting TIPE2 gene

Objective To package the recombinant lentivirus expressing TIPE2shRNA.Methods Inquiring and synthesizing seven pairs shRNA sequences targeting TIPE2,then inserted into pLKO.1lentivirus vector.After the constructions were verified to be correct,co-transfected them with pCDNA3.1/3×FLAG TIPE2plasmids into 293T cells,respectively.Then,Non target shRNA,the highest silence efficiency plasmid were co-transfected with psPAX2,pMD2.G into 293Tcells for lentivirus packaging,respectively.Results The No.4plasmid was screened by Western blotting that has the highest silence efficiency and followed by virus packing.The infection and silence efficiency was confirmed by immunofluorescence and Western blot.Conclusion The recombinant lentivirus expressing shRNA targeting TIPE2gene was packaged successfully.