Objective: To construct and identify the lentivirus vectors that interferes SET gene expression in liver cells, which would provide technical support for researching the role of SET in the mechanism of toxicity on TCE.Methods: Be based upon SET sequences retrieved through NCBI and literature reviewed, 5 pairs of shRNA were designed and synthetized, and then, which were annealed, ligated to the lentivirus pLVX-shRNA1 vector.Before plasmids were extracted, single colonies were picked and identified by PCR and sequencing.The lentivirus vector with shRNA targeting human SET mRNA were transfected into 293T cells, then the lentivirus supernatant was obtained and used for infecting L-02 cell.After selection with puromycin, the SET-deficient cells were obtained.The efficiency of gene knockdown were determined by real-time PCR and western blotting.Results: ShRNA was inserted into pLVX-shRNA1 vector, the restructured vector was transfected into 293T cells and high-titer lentivirus was formed.The lentivirus was transducted into L-02 cells, SET-deficient cells were obtained after selection.Conclusion: SET-deficient cells were successfully constructed with using lentivirus-mediated RNA interference technology.