Cationic Liposome-Mediated Transfection with Lipofectin™ Reagent

  • Philip L. Felgner
Part of the Methods in Molecular Biology book series (MIMB, volume 7)

Abstract

Since their original description, liposomes have been discussed as vehicles that could be used as carriers of pharmaceutically active agents (1), and their potential for use as carriers of genetic information has been examined (2,3). Some encouraging DNA-delivery results have been obtained; however, the methodology has had some fundamental diffkulties (4, 5, 6, 7, 8). Chief among these is that liposomes do not generally fuse with the target cell surface, but are taken up phagocytically, and the polynucleotides are subsequently subjected to the action of digestive enzymes in the lysosomal compartment. Another practical problem with conventional liposome technology results because the internal dimensions of typical liposomes may be too small to accommodate large macromolecules, such as DNA or RNA, resulting in low capturing efficiency. In addition, conventional liposomal methodology involves a relatively inconvenient multistep preparation procedure.

Keywords

Vortex Toxicity EDTA Citrate Heparin 

References

  1. 1.
    Bangham, A. D. (1981) Introduction, in Liposomesfknn Physical Structure to Therapeutic Application (Knight, G.) ed. Elsevier North-Holland, New York.Google Scholar
  2. 2.
    Straubinger, R. M. and Papahadjopoulos, D. (1983) Liposomes as carriers for intracellular delivery of nucleic acids. Methods Enzymol. 101, 512–527.PubMedCrossRefGoogle Scholar
  3. 3.
    Cudd, A, and Nicolau, C. (1984) Entrapment of recombinant DNA in liposomes and its transfer and expression of eukaryotic cells, Liposome Technology vol. II, CRC, Boca Raton, FL, pp. 207–221.Google Scholar
  4. 4.
    Mannino, R. J. and Fould-Fogerite, S. (1988) Liposome mediated gene transfer. Biotechniques6, 682–690.Google Scholar
  5. 5.
    Itani, T., Ariga, H., Yamaguchi, N., Tadakuma, T., and Yasuda, T. (1987) A simple and efficient liposome method for transfection of DNA into mammalian cells grown in suspension. Gene 56, 267–276.PubMedCrossRefGoogle Scholar
  6. 6.
    Nicolau, C., Legrand, A., and Grosse, C. E. (1987) Liposomes as carriers for in vivo gene transfer and expression. Meth. Enxymol. 149, 157–176.CrossRefGoogle Scholar
  7. 7.
    Wang, C. Y. and Huang, L. (1987) Ph-sensitive immunoliposomes mediate target cell-specific delivery and controlled expression of a foreign gene in mouse. Proc. Natl AcadSci. USA84, 7851–7855.Google Scholar
  8. 8.
    Kaneda Y, Iwai, R., and Ucheda, T. (1989) Increased expression of DNA cointroduced with nuclear protein in adult rat liver. Science 243, 375–378.PubMedCrossRefGoogle Scholar
  9. 9.
    Felgner, P. L., Gadek, T. R., Holm, M., Roman, R., Chan, H. W., Wenze, M., Northrop, J. P., Ringold, G. M., and Danielsen, M. (1987) Lipofection: A highly efficient, lipidmediated DNA-transfection procedure. Proc. NatL Acad. Sci. USA 84, 7413–7417.CrossRefGoogle Scholar
  10. 10.
    Israelachvili, J. N., Mitchell, D. J., and Ninham, B. W. (1977) Theory of self-assembly of lipid bilayers and vesicles. Biochtm. BiOphys. Acta 470, 185–201.CrossRefGoogle Scholar
  11. 11.
    Felgner, P. L. and Holm, M. (1989) Cationic liposome-mediated transfection. Focus 11:2, 21–25.Google Scholar
  12. 12.
    Duzgiines, N., Goldstein, J. A., Friend, D. S., and Felgner, P. L. (1989) Fusion of liposomes containing a novel cationic lipid N[1-(2,3-Dioleyloxy)Propyl]-N,N,N-Tri-methylammonium: Induction by multivalent anions and asymmetric fusions with acidic phospholipid vesicles. Biochemistry 28, 9179–9184.CrossRefGoogle Scholar
  13. 13.
    Felgner, P. L. and Ringold, G. M. (1989) Cationic liposome-mediated transfection. Nature 337, 387,388.CrossRefGoogle Scholar
  14. 14.
    Malone, R., Feigner, P. L., and Verma, I. (1989) Lipofectin-mediated RNA transfec-tion. A-a: Nail. Acad. Sci. USA , 6077–6081.CrossRefGoogle Scholar
  15. 15.
    Reid, T. R., Felgner, P. L., and Ringold, G. M. High efficiency RNA transfection using lipofectin: Poly IC mediated cytotoxicity in interferon treated cells (submitted to j Biol. Chem.).Google Scholar
  16. 16.
    Chen, C. A. and Okayama, H. (1988) High-efliciency transformation of mammalian cells by plasmid DNA. Biotechniques 6, 632–638.PubMedGoogle Scholar
  17. 17.
    Weiss, B., Nitschko, H., Ghattas, I., Wright, R., and Shlesinger, S. (1989) Evidence for specificity in the encapsidation of sindbis RNAs. J ViiZ. 63, 531~5318.Google Scholar
  18. 18.
    Chang, A. C. Y. and Brenner, D. G. (1988) Cationic liposome-mediated transfection: A new for the introduction of DNA into mammalii cells. Focus 10, 6669.Google Scholar
  19. 19.
    Stamatatos, L., Leventis, R., Zuckermann, M.J., and Silvius, J. R. (1988) Interactions of cationic lipid vesicles with negatively charged phospholipid vesicles and biological membranes. Biochemistry 27, 3917–3925.PubMedCrossRefGoogle Scholar

Copyright information

© The Humana Press Inc., Clifton, NJ 1991

Authors and Affiliations

  • Philip L. Felgner
    • 1
  1. 1.Voca;, Inc.San Diego

Personalised recommendations