Abstract
Following Fraley and colleagues’ initial discovery of liposomes (1), coupled with Behr’s discovery of the ability of cationic lipids to interact with and condense negatively charged DNA (2), Felgner et al. described the use of synthetic cationic lipids as a DNA delivery tool in 1987 (3). The authors reported that a positively charged lipid, specifically N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA), could form liposomes under biologically relevant conditions either alone or in combination with neutral phospholipids. More importantly, these cationic liposome vesicles were demonstrated to react spontaneously with anionic DNA to form lipid-DNA complexes, or “lipoplexes” (4). These lipoplexes demonstrated effective binding to cells in vitro and facilitation of intracellular delivery of a transgene. Felgner provides a detailed history of the progress leading up to and including lipofection (5).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Fraley, R., Subramani, S., Berg, P., and Papahadjopoulos, D. (1980) Introduction of liposome-encapsulated SV40 DNA into cells. J. Biol. Chem. 255, 10431–10435.
Behr, J. P. (1986) DNA strongly binds to micelles and vesicles containing lipopo-lyamines or lipointercalants. Tetrahedron Lett. 27, 5861–5864.
Felgner, P. L., Gadek, T. R., Holm, M., Roman, R., Chan, H. W., Wenz, M., et al. (1987) Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc. Natl. Acad. Sci. USA 84, 7413–7417.
Felgner, P. L., Barenholz, Y., Behr, J. P., Cheng, S. H., Cullis, P., Huang, L., et al. (1997) Nomenclature for synthetic gene delivery systems. Hum. Gene Ther. 8, 511–512.
Felgner, P. L. (1999) Progress in gene delivery research and development, in Nonviral Vectors for Gene Therapy (Huang, L., Hung, M.-C., and Wagner, E., eds.), Academic Press, San Diego, CA., pp. 25–38.
Chesnoy, S. and Huang, L. (2000) Structure and function of lipid-DNA complexes for gene delivery. Ann. Rev. Biophys. Biomol. Struct. 29, 27–47.
Pedroso de Lima, M. C., Simoes, S., Pires, P., Faneca, H., and Duzgunes, N. (2001) Cationic lipid-DNA complexes in gene delivery: from biophysics to biological applications. Adv. Drug Deliv. Rev. 47, 277–294.
James, M. B. and Giorgio, T. D. (2000) Nuclear-associated plasmid, but not cell-associated plasmid, is correlated with transgene expression in cultured mammalian cells. Mol. Ther. 1, 339–346.
Lechardeur, D., Sohn, K. J., Haardt, M., Joshi, P. B., Monck, M., Graham, R. W., et al. (1999) Metabolic instability of plasmid DNA in the cytosol: a potential barrier to gene transfer. Gene Ther. 6, 482–497.
Tseng, W. C., Haselton, F. R., and Giorgio, T. D. (1997) Transfection by cationic liposomes using simultaneous single cell measurements of plasmid delivery and transgene expression. J. Biol. Chem. 272, 25641–25647.
Gustafsson, J., Arvidson, G., Karlsson, G., and Almgren, M. (1995) Complexes between cationic liposomes and DNA visualized by cryo-TEM. Biochim. Biophys. Acta 1235, 305–312.
Sternberg, B., Sorgi, F. L., and Huang, L. (1994) New structures in complex formation between DNA and cationic liposomes visualized by freeze-fracture electron microscopy. FEBS Lett. 356, 361–366.
Templeton, N. S., Lasic, D. D., Frederik, P. M., Strey, H. H., Roberts, D. D., and Pavlakis, G. N. (1997) Improved DNA: liposome complexes for increased systemic delivery and gene expression. Nat. Biotechnol. 15, 647–652.
Ross, P. C. and Hui, S. W. (1999) Lipoplex size is a major determinant of in vitro lipofection efficiency. Gene Ther. 6, 651–659.
van der Woude, I., Visser, H. W., ter Beest, M. B., Wagenaar, A., Ruiters, M. H., Engberts, J. B., and Hoekstra, D. (1995) Parameters influencing the introduction of plasmid DNA into cells by the use of synthetic amphiphiles as a carrier system. Biochim. Biophys. Acta 1240, 34–40.
Zabner, J., Fasbender, A. J., Moninger, T., Poellinger, K. A., and Welsh, M. J. (1995) Cellular and molecular barriers to gene transfer by a cationic lipid. J. Biol. Chem. 270, 18997–19007.
Coonrod, A., Li, F. Q., and Horwitz, M. (1997) On the mechanism of DNA trans-fection: efficient gene transfer without viruses. Gene Ther. 4, 1313–1321.
Zhou, X. and Huang, L. (1994) DNA transfection mediated by cationic liposomes containing lipopolylysine: characterization and mechanism of action. Biochim. Biophys. Acta 1189, 195–203.
Friend, D. S., Papahadjopoulos, D., and Debs, R. J. (1996) Endocytosis and intra-cellular processing accompanying transfection mediated by cationic liposomes. Biochim. Biophys. Acta 1278, 41–50.
Hui, S. W., Langner, M., Zhao, Y. L., Ross, P., Hurley, E., and Chan, K. (1996) The role of helper lipids in cationic liposome-mediated gene transfer. Biophys. J. 71, 590–599.
Farhood, H., Serbina, N., and Huang, L. (1995) The role of dioleoyl phosphatidylethanolamine in cationic liposome mediated gene transfer. Biochim. Biophys. Acta 1235, 289–295.
Simoes, S., Slepushkin, V., Duzgunes, N., and Pedroso de Lima, M. C. (2001) On the mechanisms of internalization and intracellular delivery mediated by pH-sensitive liposomes. Biochim. Biophys. Acta 1515, 23–37.
Invitrogen and Life Technologies (1999) Guide to Eukaryotic Transfections with Cationic Lipid Reagents. Rockville, MD, pp. 1–33.
Ciccarone, V. and Hawley-Nelson, P. (1995) Lipofectin transfection activity increased by protocol improvement. Focus 17(3), 103.
Freshney, R. I. (1993) Culture of Animal Cells. Alan R. Liss, Inc., New York
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2004 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Wyatt, S.K., Giorgio, T.D. (2004). DNA Delivery to Cells in Culture Using Cationic Liposomes. In: Heiser, W.C. (eds) Gene Delivery to Mammalian Cells. Methods in Molecular Biology™, vol 245. Humana Press. https://doi.org/10.1385/1-59259-649-5:83
Download citation
DOI: https://doi.org/10.1385/1-59259-649-5:83
Publisher Name: Humana Press
Print ISBN: 978-1-58829-086-1
Online ISBN: 978-1-59259-649-2
eBook Packages: Springer Protocols