Construction of KIR2DS2 lentiviral vector

Ojbective: To construct a lentiviral vector of RNA interference（RNAi） of KIR2DS2 gene.Methods: The effective sequence of siRNA targeting KIR2DS2 gene was confirmed in our study.The complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pSicoR-GFP vector,which contained U6 promoter and green fluorescent protein（GFP）.The resulting lentiviral vector containing KIR2DS2 shRNA was named LV-sh KIR2DS2,and it was confirmed by PCR and sequencing.293T cells were cotransfected with lentiviral vector LV-sh KIR2DS2 and packaging system.All virus stocks were produced by Lipofectamine 2000-mediated transfection.The titer of virus was tested according to the expression level of GFP.Results: PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of KIR2DS2 was constructed successfully.The titer of virus tested according to the expression level of GFP was 6×10⁸ TU/ml.Conclusions: The lentivirus RNAi vector of KIR2DS2 is constructed successfully.