TITLE:
The Mechanism of Lipofectamine 2000 Mediated Transmembrane Gene Delivery
AUTHORS:
Shaohui Cui, Shubiao Zhang, Huiying Chen, Bing Wang, Yinan Zhao, Defu Zhi
KEYWORDS:
Gene Delivery; Cationic Liposomes; Transfection Efficiency; Endocytic Pathway; Inhibitor
JOURNAL NAME:
Engineering,
Vol.4 No.10B,
October
21,
2013
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.