The Mechanism of Lipofectamine 2000 Mediated Transmembrane Gene Delivery

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.

Share and Cite:

S. Cui, S. Zhang, H. Chen, B. Wang, Y. Zhao and D. Zhi, "The Mechanism of Lipofectamine 2000 Mediated Transmembrane Gene Delivery," Engineering, Vol. 4 No. 10B, 2012, pp. 172-175. doi: 10.4236/eng.2012.410B045.

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1] K. A. High. Gene therapy-the moving finger. Nature 435: 577-579, 2005.
[2] Y. Zhang, H. M. Li and S. Jing , DC-Chol/DOPE cationic liposomes: A comparative study of the influence factors on plasmid pDNA and siRNA gene delivery. International Journal of Pharmaceutics May 10; 390(2):198-207, 2010.
[3] J. Rejman, A. Bragonzi and M. Conese, Role of clathrin- and caveolae mediated endocytosis in gene transfer mediated by lipo- and polyplexes. Mol Ther 2005;12:468-74,2005.
[4] M. Satyajit and E. P. Richard, Pathways of clath-rin-independent endocytosis. Nat Rev 8:603-12,2007.
[5] K. Sandvig , S. Pust and T. Skotland, Clathrin-independent endocytosis: mechanisms and function. Curr Opin Cell Biol.Aug;23(4):413-20, 2011.
[6] M. T. Howes, S. Mayor and R. G. Parton, Molecules, mechanisms, and cellular roles of clathrin-independent endocytosis. Curr Opin Cell Biol 22(4): 519-27, 2010.
[7] B. Emmanuelle, B. Sandrine and G. Lucie, Hepatitis C Virus Entry Depends on Clathrin-Mediated Endocytosis. J Virol. 80(14):6964–6972, July 2006.
[8] D. Hoekstra, J. Rejman and L. Wasungu, Gene delivery by cationic lipids: in and out of an endosome.Biochem.Soc Trans. Feb;35(Pt 1):68-71,2007.
[9] Y. Obata, G. Ciofani and V. Raffa, Evaluation of cationic liposomes composed of an amino acid–based of an endosome. Biochemical Society Transactions 35: 68-71,2010.

Copyright © 2024 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.