The Mechanism of Lipofectamine 2000 Mediated Transmembrane Gene Delivery

Abstract

Inthis paper, the relatived mechanism between lipofectamine 2000 mediatedtransmembrane gene delivery and endocytic pathway were investigated. Clathrin and caveolae-mediated endocytic pathwaycontributions to transfection efficiency were studied. The inhibitorsof endocytosis were used to treat HEp-2 cells before lipofectamine 2000/pGFP-N2transfection. Transfection efficiency was evaluated with green fluorescence protein(GFP) expression assays. Cell viability and cytotoxicity were evaluated withMTT method. The results indicated that inhibitors of clathrin (chlorpromazineor wortmannin) and caveolin (genistein) could reduce the cell transfection efficiency observably.Both clathrin and caveolae-mediated endocytic pathways play important roles intransmembrane gene delivery.