The Mechanism of Lipofectamine 2000 Mediated Transmembrane Gene Delivery

Shaohui Cui, Shubiao Zhang, Huiying Chen, Bing Wang, Yinan Zhao, Defu Zhi

Key Laboratory of Bio-chemistry Engineering - The State Ethnic Affairs Commission-Ministry of Education, Dalian Nationalities University, Dalian, China.

DOI: 10.4236/eng.2012.410B045                

Abstract

In this paper, the relatived mechanism between lipofectamine 2000 mediated transmembrane gene delivery and endocytic pathway were investigated. Clathrin and caveolae-mediated endocytic pathway contributions to transfection efficiency were studied. The inhibitors of endocytosis were used to treat HEp-2 cells before lipofectamine 2000/pGFP-N2 transfection. Transfection efficiency was evaluated with green fluorescence protein (GFP) expression assays. Cell viability and cytotoxicity were evaluated with MTT method. The results indicated that inhibitors of clathrin (chlorpromazine or wortmannin) and caveolin (genistein) could reduce the cell transfection efficiency observably. Both clathrin and caveolae-mediated endocytic pathways play important roles in transmembrane gene delivery.

Keywords

Gene Delivery; Cationic Liposomes; Transfection Efficiency; Endocytic Pathway; Inhibitor

Conflicts of Interest

The authors declare no conflicts of interest.

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