Construction and Identification of Plasmid Mediated RNA Interference Targeting Egr-1

Objective:To construct and efficient screening out the RNA interference (RNAi) plasmid for the early growth response gene-1 (Egr-1). Methods:In accordance with mRNA sequence of gene Egr-1, 3 oligonucleotide sequence with small hairpin structure were cloned into empty carrier pGCSIL-GFP to construct recombinant plasmid, meanwhile, the plasmid not directing towards any differentia gene was designed as negative comparison. Used shRNA expressed plasmid to transfect HEK293 cells. Identified the interference efficiency of shRNA expressed plasmid to Egr-1, by observing GFP expression quantity, fluorescence quantitative PCR and identifing the expression of gene Egr-1 using quantitative western blotting. Results:3 sequence of RNAi to mouse gene Egr-1, one sequence had the interference efficiency greater than 70%. Conclusion:Successfully constructed a RNAi highly active expression plasmid to mouse gene Egr-1.