Abstract
The use of cationic lipids or cationic polymers to mediate the transfer of nucleic acids into mammalian cells has become a widely applied technology in recent years. The principal reasons for this have been the ease with which the methodology can be applied to a wide range of cell types; the relatively low cytotoxicity compared to other techniques; the high efficiency of nucleic acid transfer in comparison with methods such as calcium phosphate or diethylaminoethyl-dextran-mediated transfection; and the potential application of these systems to human gene therapy. The use of positively charged lipid-based macromolecules to deliver nucleic acids makes use of the fact that DNA, RNA, and oligonucleotides carry a negative charge caused by the phosphate groups that form the backbone of these molecules. The electrostatic interaction between the negatively charged nucleic acid and the positively charged macromolecule induces a range of structural changes that vary, depending on the macro-molecule used. In general, however, the process results in condensation or compaction of the nucleic acid and physical association of the nucleic acid with the lipid. The interaction generates a complex that is more amenable to cellular uptake, protects sufficient nucleic acid molecules to allow trafficking to the nucleus, and, in at least some cases, may also facilitate transfer into the nucleus.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Capaccioli, S., Pasquale, G. D., Mini, E., Mazzei, T., and Quattrone, A. (1993) Cationic lipids improve antisense oligonucleotide uptake and prevent degradation in cultured cells and in human serum. Biochem. Biophys. Res. Comm. 197, 818–825.
Lewis, J. G., Lin, K.-Y., Kothavale, A., Flanagan, W. M., Matteucci, M. D., DePrince, R. B., et al. (1996) Serum-resistant cytofectin for delivery of antisense oligonucleotides and plasmid DNA. Proc. Natl. Acad. Sci. USA 93, 3176–3181.
Kronenwett, R., Steidi, U., Kirsch, M., Sczakiel, G., and Haas, R. (1998) Oligodeoxyribonucleotide uptake in primary human hematopoietic cells is enhanced by cationic lipids and depends on the hematopoietic cell subset. Blood 91, 852–862.
Chen, M., Compton, S. T., Coviello, V. F., Green, E. D., and Ashlock, M. A. (1997) Transient gene expression from yeast artificial chromosome DNA in mammalian cells is enhanced by adenovirus. Nucleic Acids Res. 25, 4416–4418.
Ikeno, M., Grimes, B., Okazaki, T., Nakano, M., Saitoh, K., Hoshino, H., et al. (1998) Construction of YAC-based mammalian artificial chromosomes. Nature Biotechnol. 16, 431–439.
Lu, D., Benjamin, R., Kim, M., Conry, R. M., and Curiel, D. T. (1994) Optimization of methods to achieve mRNA-mediated transfection of tumor cells in vitro and in vivo employing cationic liposome vectors. Cancer Gene Ther. 1, 245–252.
Kariko, K., Kuo, A., Barnathan, E. S., and Langer, D. J. (1998) Phosphateenhanced transfection of cationic lipid-complexed mRNA and plasmid DNA. Biochim. Biophys. Acta 1369, 320–334.
Cole-Strauss, A., Yoon, K., Xiang, Y., Byrne, B. C., Rice, M. C., Gryn, J., Holloman, W. K., and Kmiec, E. B. (1996) Correction of the mutation responsible for sickle cell anemia by an RNA-DNA oligonucleotide. Science 273, 1386–1389.
Kren, B. T., Bandyopadhyay, P., and Steer, C. J. (1998) In vivo site-directed mutagenesis of the factor IX gene by chimeric RNA/DNA oligonucleotides. Nature Med. 4, 285–290.
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., 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.
Gao, X. and Huang, L. (1995) Cationic liposome-mediated gene transfer. Gene Ther. 2, 710–722.
Gao, X. (1997) Cationic lipid-based gene delivery: an update, in Gene Therapy for Diseases of the Lung (Brigham, K. L., ed.), Marcel Dekker, New York, pp. 99–112.
Gao, X. and Huang, L. (1991) Novel cationic liposome reagent for efficient transfection of mammalian cells. Biochem. Biophys. Res. Comm. 179, 280–285.
Felgner, J. H., Kumar, R., Sridhar, C. N., Wheeler, C. J., Tsai, Y. J., Border, R., et al. (1994) Enhanced gene delivery and mechanism studies with a novel series of cationic lipid formulations. J. Biol. Chem. 269, 2550–2561.
Lee, E. R., Marshall, J., Siegel, C. S., Jiang, C., Yew, N. S., Nichols, M. R., et al. (1996) Detailed analysis of structures and formulations of cationic lipids for efficient gene transfer to the lung. Hum. Gene Ther. 7, 1701–1717.
Wheeler, C., Felgner, P. L., Tsai, Y. J., Marshall, J., Sukhu, L., Doh, S. G., et al. (1996) Novel cationic lipid greatly enhances plasmid DNA delivery and expression in mouse lung. Proc. Natl. Acad. Sci. USA 93, 11,454–11,459.
Gershon, H., Ghirlando, R., Guttman, S. B., and Minsky, A. (1993) Mode of formation and structural features of DNA-cationic liposome complexes used for transfection. Biochem. 32, 7143–7151.
Eastman, S., Siegel, C., Tousignant, J., Smith, A. E., Cheng, S. H., and Scheule, R. K. (1997) Biophysical characterization of cationic lipid: DNA complexes. Biochim. Biophys. Acta 1325, 41–62.
Ferrari, M. E., Nguyan, C. M., Zelphati, O., Tsai, Y., and Felgner, P. L. (1998) Analytical methods for the characterization of cationic lipid-nucleic acid complexes. Hum. Gene Ther. 9, 341–351.
Felgner, P. L. and Rinegold, G. M. (1989) Cationic liposome mediated transfection. Nature 337, 387–388.
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.
Rädler, J., Koltover, I., Salditt, T., and Safinya, C. R. (1997) Structure of DNAcationic liposome complexes: DNA intercalation in multilamellar membranes in distinct interhelical packing regimes. Science 275, 810–81
Spector, M. S. and Schnur, J. M. (1997) DNA ordering on a lipid membrane. Science 275, 791–792.
Smith, J. G., Walzem, R. L., and German, J. B. (1993) Liposomes as agents of DNA transfer. Biochim. Biophys. Acta 1154, 327–340.
Legendre, J. Y. and Szoka, F. C. (1992) Delivery of plasmid DNA into mammalian cell lines using pH sensitive liposomes: comparison with cationic liposomes. Pharm. Res. 9, 1235–1242.
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.
Wrobel, I. and Collins, D. (1995) Fusion of cationic liposomes with mammalian cells occurs after endocytosis. Biochim. Biophys. Acta 1235, 296–304.
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, 18,997–19,007.
Matsui, H., Johnson, L. G., Randell, S. H., and Boucher, R. C. (1997) Loss of binding and entry of liposome-DNA complexes decreases transfection efficiency in differentiated airway epithelial cells. J. Biol. Chem. 272, 1117–1126.
Caplen, N. J., Kinrade, E., Sorgi, F., Gao, X., Gruenert, D., Geddes, D., et al. (1995) In vitro liposome-mediated DNA transfection of epithelial cell lines using the cationic liposome DC-Chol/DOPE. Gene Ther. 2, 603–613.
Fasbender, A. J., Zabner, J., and Welsh, M. J. (1995) Optimization of cationic lipid-mediated gene transfer to airway epithelia. Am. J. Physiol. 269 (Lung Cell. Mol. Physiol.), L45–L51.
Eastman, S. J., Tousignant, J. D., Lukason, M. J., Murray, H., Siegel, C. S., Constantino, P., et al. (1997) Optimization of formulations and conditions for the aerosol delivery of functional cationic lipid:DNA complexes. Hum. Gene Ther. 8, 313–322.
Liu, Y., Mounkes, L. C., Liggitt, H. D., Brown, C. S., Solodin, I., Heath, T. D., and Debs, R. J. (1997) Factors influencing the efficiency of cationic liposomemediated intravenous gene delivery. Nature Biotechnol. 15, 167–173.
Caplen, N. J., Gao, X., Hayes, P., Elaswarapu, R., Fisher, G., Kinrade, E., et al. (1994) Gene therapy for cystic fibrosis in humans by liposome-mediated DNA transfer: the production of resources and the regulatory process. Gene Ther. 1, 139–147.
Cotten, M., Baker, A., Saltik, M., Wagner, E., and Buschle, M. (1994) Lipopolysaccharide is a frequent contaminant of plasmid DNA preparations and can be toxic to primary human cells in the presence of adenovirus. Gene Ther. 1, 239–246.
Cotten, M. and Saltik, M. (1997) Intracellular delivery of lipopolysaccharide during DNA transfection activates a lipid A-dependent cell death response that can be prevented by polymyxin B. Hum. Gene Ther. 8, 555–561.
Behr, J. P., Demeneix, B., Loeffler, J. P., and Mutul, J. P. (1989) Efficient gene transfer into mammalian primary endocrine cells with lipopolyamine-coated DNA. Proc. Natl. Acad. Sci. USA 86, 6982–6986.
Lasic, D. D. (1997) Liposomes in Gene Delivery. CRC, Boca Raton, FL.
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2000 Humana Press Inc.
About this protocol
Cite this protocol
Caplen, N.J. (2000). Nucleic Acid Transfer Using Cationic Lipids. In: Kmiec, E.B. (eds) Gene Targeting Protocols. Methods in Molecular Biology™, vol 133. Humana Press. https://doi.org/10.1385/1-59259-215-5:1
Download citation
DOI: https://doi.org/10.1385/1-59259-215-5:1
Publisher Name: Humana Press
Print ISBN: 978-0-89603-360-3
Online ISBN: 978-1-59259-215-9
eBook Packages: Springer Protocols