Construction and Identification of the RNA Interference Vector Targeting Sp1 Gene

Objective:To construct the RNA interference(RNAi) vector targeting Sp1 gene.Methods:According to Sp1 cDNA coding sequence,the specific RNAi fragment targeting Sp1 gene was designed and synthesized,which was cloned into pSilencer 3.1-H1 neo plasmid vector,and the small interfering RNA(siRNA) eukaryotic expression vector pSilencer 3.1-Sp1 targeting Sp1 gene was constructed.The pSilencer 3.1 negtive vector and pSilencer 3.1-Sp1 vector were transfected respectively into HeLa cells by LipofectAMINE2000.RT-PCR and Western blot methods were carried out for the identification of integration of RNAi vectors on mRNA and protein levels.Results:The specific siRNA eukaryotic expression vector pSilencer 3.1-Sp1 targeting Sp1 gene was constructed successfully,and the knockdown effects on mRNA and protein levels were confirmed.Conclusion:Sp1 gene expression can be suppressed markedly by specific siRNA in HeLa cells.Based on the results,the further studying on the biological functions and mechanisms of Sp1 will be carried out.