Skip to main content

DNA Delivery to Cells in Culture Using Cationic Liposomes

  • Protocol
Book cover Gene Delivery to Mammalian Cells

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 245))

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).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Fraley, R., Subramani, S., Berg, P., and Papahadjopoulos, D. (1980) Introduction of liposome-encapsulated SV40 DNA into cells. J. Biol. Chem. 255, 10431–10435.

    PubMed  CAS  Google Scholar 

  2. Behr, J. P. (1986) DNA strongly binds to micelles and vesicles containing lipopo-lyamines or lipointercalants. Tetrahedron Lett. 27, 5861–5864.

    Article  CAS  Google Scholar 

  3. 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.

    Article  PubMed  CAS  Google Scholar 

  4. 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.

    Article  PubMed  CAS  Google Scholar 

  5. 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.

    Chapter  Google Scholar 

  6. Chesnoy, S. and Huang, L. (2000) Structure and function of lipid-DNA complexes for gene delivery. Ann. Rev. Biophys. Biomol. Struct. 29, 27–47.

    Article  CAS  Google Scholar 

  7. 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.

    Google Scholar 

  8. 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.

    Article  PubMed  CAS  Google Scholar 

  9. 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.

    Article  PubMed  CAS  Google Scholar 

  10. 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.

    Article  PubMed  CAS  Google Scholar 

  11. 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.

    Article  PubMed  Google Scholar 

  12. 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.

    Article  PubMed  CAS  Google Scholar 

  13. 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.

    Article  PubMed  CAS  Google Scholar 

  14. Ross, P. C. and Hui, S. W. (1999) Lipoplex size is a major determinant of in vitro lipofection efficiency. Gene Ther. 6, 651–659.

    Article  PubMed  CAS  Google Scholar 

  15. 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.

    Article  PubMed  Google Scholar 

  16. 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.

    Article  PubMed  CAS  Google Scholar 

  17. 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.

    Article  PubMed  CAS  Google Scholar 

  18. 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.

    Article  PubMed  CAS  Google Scholar 

  19. 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.

    Article  PubMed  Google Scholar 

  20. 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.

    Article  PubMed  CAS  Google Scholar 

  21. 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.

    Article  PubMed  Google Scholar 

  22. 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.

    Google Scholar 

  23. Invitrogen and Life Technologies (1999) Guide to Eukaryotic Transfections with Cationic Lipid Reagents. Rockville, MD, pp. 1–33.

    Google Scholar 

  24. Ciccarone, V. and Hawley-Nelson, P. (1995) Lipofectin transfection activity increased by protocol improvement. Focus 17(3), 103.

    Google Scholar 

  25. Freshney, R. I. (1993) Culture of Animal Cells. Alan R. Liss, Inc., New York

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints 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

Publish with us

Policies and ethics