Gene Therapy pp 185-194 | Cite as

Lipid Gene Trasfer, a Story of Simplicity and Complexity

  • Natasha J. Caplen
Conference paper
Part of the NATO ASI Series book series (volume 105)


It is 10 years since the first description of the use of a positively charged lipid to deliver nucleic acids to mammalian cells (Feigner et al., 1987). In essence the methodology of liposome-mediated transfer of nucleic acids is very simple. Cells are exposed to a complex of nucleic acid and lipid formed by simply mixing the two components together. The lipid in some way facilities both uptake of the nucleic acid by, and transport within a cell, usually enhancing transfer many fold over that seen if nucleic acid alone is used. However, this description is proving to be something of an over simplification of what is probably a much more complex process. This review will discuss the basic chemistry of the most widely used cationic liposomes, current models for DNA-liposome complexes, the chemical and biological factors that effect the efficiency of DNA transfer, and the proposed trafficking mechanisms that facilitate DNA transfer to the nucleus.


Cationic liposomes liposome-mediated gene transfer 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Alton, E.W.F.W., Middleton, P.G., Caplen, N.J., Smith, S.N., Steel, D.M., Munkonge, F.M., Jeffery, P.K., Geddes, D.M., Hart, S.L., Williamson, R., Fasold, K.I., Miller, A.D., Dickinson, P., Stevenson, B.J., McLachlan, G., Dorin, J.R., and Porteous, D.J. (1993) Non-invasive liposome-mediated gene delivery can correct the ion transport defect in cystic fibrosis mutant mice. Nature Genetics 5, 135–142.PubMedCrossRefGoogle Scholar
  2. 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.CrossRefGoogle Scholar
  3. Canonico, A.E., Conary, J.T., Meyrick, B.O., and Brigham, K.L. (1994) Aerosol and intravenous transfection of human αl-antitrypsin gene to lungs of rabbits. Am. J. Respir. Cell Mol. Biol. 10, 24–29.PubMedGoogle Scholar
  4. Caplen, N.J., Kinrade, E., Sorgi, F., Gao, X., Gruenert, D., Geddes, D., Coutelle, C., Huang, L., Alton, E.W.F.W., and Williamson, R. (1995a) In vitro liposome- mediated DNA transfection of epithelial cell lines using the cationic liposome DC- Chol/DOPE. Gene Therapy 2, 603–613.PubMedGoogle Scholar
  5. Caplen, N.J., Alton, E.W.F.W., Middleton, P.G., Dorin, J.R., Stevenson, B.J., Gao, X., Durham, S., Jeffery, P.K., Hodson, M.E., Coutelle, C., Huang, L., Porteous, D.J., Williamson, R., and Geddes, D.M. (1995b) Liposome-mediated CFTR gene transfer to the nasal epithelium of patients with cystic fibrosis. Nature Medicine 1, 39–46. Addendum 1 (3) 272.PubMedCrossRefGoogle Scholar
  6. Eastman, S.J., Tousignant, J.D., Lukason, M.J., Murray, H., Siegel, C.S., Constantino, P., Harris, D.J., Cheng, S.H., and Scheule, R.K. (1997) Optimization of formulations and conditions for the aerosol delivery of functionla cationic lipid:DNA complexes. Human Gene Therapy 8, 313–322.PubMedCrossRefGoogle Scholar
  7. Fasbender, A.J., Zabner, J., and Welsh, M.J. (1995) Optimization of cationc lipid-mediated gene transfer to airway epithelia. Am. J. Physiol. 269 (Lung Cell. Mol. Physiol.), L45-L51.PubMedGoogle Scholar
  8. Feigner, P.L., Gadek, T.R., Holm, M., Roman, R., Chan, H.W., Wenz, M., Northrop, J.P., Ringolg, G.M., and Danielsen, M. (1987) Lipofection: A highly efficient, lipid-mediated DNA-transfection procedure. Proc. Natl. Acad. Sci. (USA) 84, 7413–7417.CrossRefGoogle Scholar
  9. Feigner, P.L. and Rinegold, G.M. (1989) Cationic liposome mediate transfection. Nature 337, 387–388.CrossRefGoogle Scholar
  10. Feigner, J.H., Kumar, R., Sridhar, C.N., Wheeler, C.J., Tsai, Y.J., Border, R., Ramsey, P., Martin, M., and Feigner, P.L. (1994) Enhanced gene delivery and mechanism studies with a novel series of cationic lipid formulations. Jour. Biol. Chem. 269, 2550–2561.Google Scholar
  11. Feigner, P.L., Barenholz, Y., Behr, J.P., Cheng, S.H., Cullis, P., Huang, L., Jessee, J.A., Seymour, L., Szoka, F., Thierry, A.R., Wagner, E., and Wu, G. (1997) Nomenclature for synthetic gene delivery systems. Human Gene Therapy 8, 511–512.CrossRefGoogle Scholar
  12. Gao, X. and Huang, L. (1991) A novel cationic liposome reagent for efficient transfection of mammalian cells. Biochem. Biophys. Res. Comm. 179, 280–285.PubMedCrossRefGoogle Scholar
  13. 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. Biochemistry 32, 7143–7151.PubMedCrossRefGoogle Scholar
  14. Gill, D., Southern, K.W., Mofford, K.A., Seddon, T., Huang, L., Sorgi, F., Thomson, A., MacVinish, L.J., Ratcliff, R., Bilton, D., Lane, D.J., Littlewood, J.M., Webb, A.K., Middleton, P.G., Colledge, W.H., Cuthbert, A.W., Evans, M.J., Higgins, C.F., and Hyde, S.C. (1997) A placebo-controlled study of liposome-mediated gene transfer to the nasal epithelium of patients with cystic fibrosis. Gene Therapy 4, 199–209.PubMedCrossRefGoogle Scholar
  15. 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.PubMedCrossRefGoogle Scholar
  16. Hyde, S., Gill, D., Higgins, C., Treizie, A., MacVinish, L.J., Cuthburt, A., Ratclife, R., Evans, M., and Colledge, W. (1993) Correction of the ion transport defect in cystic fibrosis transgenic mice by gene therapy. Nature 362, 250–255.PubMedCrossRefGoogle Scholar
  17. Lee, E.R., Marshall, J., Siegel, C.S., Jiang, C., Yew, N.S., Nichols, M.R., Nietupski, J.B., Ziegler, R.J., Lane, M.B., Wang, K.X., Wan, N.C., Scheule, R.K., Harris, D.J., Smith, A.E., and Cheng, S.H. (1996) Detailed analysis of structures and formulations of cationic lipids for efficient gene transfer to the lung. Human Gene Therapy 7, 1701–1717.PubMedCrossRefGoogle Scholar
  18. Legendre, J.Y. and Szoka, F.C. (1992) Delivery of plamsmid DNA into mammalian cell lines using pH senstive liposomes: comparison with cationic liposomes. Pharm. Res. 9, 1235–1242.PubMedCrossRefGoogle Scholar
  19. 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 liposome-mediated intravenous gene delivery. Nature Biotechnology 15, 167–173.PubMedCrossRefGoogle Scholar
  20. Marcel, T. and Grausz, J.D. (1997) The TMC worldwide gene therapy enrollment report, end 1996. Human Gene Therapy 8, 775–800.PubMedCrossRefGoogle Scholar
  21. Matsui, H., Johnson, L.G., Randeil, S.H., and Boucher, R.C. (1997) Loss of binding and entry of liposome-DNA complexes decreases trasnfection efficiency in differentiated airway epithelial cells. Jour. Biol. Chem. 272, 1117–1126.CrossRefGoogle Scholar
  22. McLachlan, G., Davidson, D.J., Stevenson, B.J., P.Dickinson, Davidson-Smith, H., Dorin, J.R., and Porteous, D.J. (1995) Evaluation in vitro and in vivo of cationic liposome-expression construct complexes for cystic fibrosis gene therapy. Gene Therapy 2, 614–622.PubMedGoogle Scholar
  23. Nabel, G.J., Nabel, E.G., Yang, Z.-Y., Fox, B.A., Plutz, G.E., Gao, X., Huang, L., Shu, S., Gordon, D., and Chang, A.E. (1993) Direct gene transfer with DNA- liposome complexes in melanoma: Expression, biologic activity, and lack of toxicity in humans. Proc. Natl. Acad. Sci. (USA) 90, 11307–11311.CrossRefGoogle Scholar
  24. Nabel, G.J., Gordon, D., Bishop, D.K., J. Nickoloff, B., Yang, Z.-Y., Araga, A., Cameron, M.J., Nabel, E.G., and Chang, A.E. (1996) Immune response in human melanoma after transfer of an allogenic class I major histocompatibility complex gene with DNA-liposome complexes. Proc. Natl. Acad. Sci. (USA)93, 15388–15393.CrossRefGoogle Scholar
  25. Plautz, G.E., Yang, Z.Y., Wu, B., Gao, X., Huang, L., and Nabel, G.J. (1993) Immunotherapy of malignacy by in vivo gene transfer into tumors. Proc. Natl. Acad. Sci. (USA) 90, 4645–4649.CrossRefGoogle Scholar
  26. Porteous, D., Dorin, J.R., McLachlan, G., Davidson-Smith, H., Davidson, H., Stevenson, B.J., Carothers, A.D., Wallace, W.A., Moralee, S., Hoenes, C., Kallmeyer, G., Michaelis, U., Naujoks, K., Ho, L.P., Samways, J.M., Imrie, M., Greening, A.P., and Innes, J.A. (1997) Evidence for safety and efficacy of DOTAP cationic liposome-mediated CFTR gene transfer to the nasal epithelium of patients with cystic fibrosis. Gene Therapy 4, 210–218.PubMedCrossRefGoogle Scholar
  27. Rädler, J., Koltover, I., Salditt, T., and Safinya, C.R. (1997) Structure of DNA- cationic liposome complexes: DNA intercalation in multilamellar membranes in distinct interhelical packing regimes. Science 275, 810–814.PubMedCrossRefGoogle Scholar
  28. Roessler, B.J. and Davidson, B.L. (1994) Direct plasmid mediated transfection of adult murine brain cells in vivo using cationic liposomes. Neurosci. Lett. 167, 5–10.PubMedCrossRefGoogle Scholar
  29. Smith, J.G., Walzem, R.L., and German, J.B. (1993) Liposomes as agents of DNA transfer. Biochim. Biophys. Acta 1154, 327–340.PubMedGoogle Scholar
  30. Smyth Templeton, N., 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. Nature Biotechnology 15, 647–652.PubMedCrossRefGoogle Scholar
  31. Spector, M.S. and Schnur, J.M. (1997) DNA ordering on a lipid membrane. Science 275, 791–792.PubMedCrossRefGoogle Scholar
  32. Stephan, D.J., Yang, Z.-Y., San, H., Simari, R.D., Wheeler, C.J., Feigner, P.L., Gordon, D., Nabel, G.J., and Nabel, E.G. (1996) A new cationic liposome DNA complex enhances the efficiency of arterial gene transfer in vivo. Human Gene Therapy 7, 1803–1812.PubMedCrossRefGoogle Scholar
  33. Sternberg, B., Sorgi, F.L., and Huang, L. (1994) New structures in complex formation between DNA and cationic liposomes visualised by freeze-fracture electron microscopy. FEBS Letters 356, 361–366.PubMedCrossRefGoogle Scholar
  34. Stewart, M.J., Plautz, G.E., Del Buono, L., Yang, Z.Y., Xu, L., Gao, X., Huang, L., Nabel, E.G., and Nabel, G.J. (1992) Gene transfer in vivo with DNA-liposome complexes: safety and acute toxicity in mice. Human Gene Therapy 3, 267–275.PubMedCrossRefGoogle Scholar
  35. Stribling, R., Brunette, E., Liggitt, D., Gaensler, K., and Debs, R. (1992) Aerosol gene delivery in vivo. Proc. Natl. Acad. Sci. (USA) 89, 11277–11281.CrossRefGoogle Scholar
  36. Thierry, A.R., Lunardi-Iskander, Y., Bryant, J.L., Rabinovich, P., Gallo, R.C., and Mahans, L.C. (1995) Systemic gene therapy: biodistribution and long-term expression of a transgene in mice. Proc. Natl. Acad. Sci. (USA) 92, 9742–9746.CrossRefGoogle Scholar
  37. Wheeler, C., Feigner, P.L., Tsai, Y.J., Marshall, J., Sukhu, L., Doh, S.G., Hartikka, J., Nietupski, J., Manthorpe, M., Nichols, M., Plewe, M., Liang, X., Norman, J., Smith, A., and Cheng, S.H. (1996) A novel cationic lipid greatly enhances plasmid DNA delivery and expression in mouse lung. Proc. Natl. Acad. Sci. (USA) 93, 11454–11459.CrossRefGoogle Scholar
  38. Wrobel, I. and Collins, D. (1995) Fusion of cationic liposomes with mammalian cells occurs after endoeytosis. Biochim. Biophys. Acta 1235, 296–304.PubMedCrossRefGoogle Scholar
  39. Xing, X., Liu, V., Xia, W., Stephens, L.C., Huang, L., Lopez-Berestein, G., and Hung, M.-C. (1997) Safety studies of the intraperitoneal injection of E1A- liposome complex in mice. Gene Therapy 4, 238–243.PubMedCrossRefGoogle Scholar
  40. Xu, Y. and Szoka, F.C. (1996) Mechanism of DNA release from cationic liposome/DNA complexes used in cell transfection. Biochemistry 35, 5616–5623.PubMedCrossRefGoogle Scholar
  41. Yoshimura, K., Rosenfield, M.A., Nakamura, H., Scherer, E.M., Pavirani, A., Lecocq, J.-P., and Crystal, R.G. (1992) Expression of the human cystic fibrosis transmembrane conductance regulator gene in the mouse lung after in vivo intratracheal plasmid-mediated gene transfer. Nuc. Acids Res. 20, 3233–3240.CrossRefGoogle Scholar
  42. 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. Jour. Biol. Chem. 270, 18997–19007.CrossRefGoogle Scholar
  43. Zhou, X., Kilbanov, A.L., and Huang, L. (1991) Lipophilic polylysines mediate efficient DNA transfection in mammalian cells. Biochim. Biophys. Acta 1065, 8–14.PubMedCrossRefGoogle Scholar
  44. Zhou, X. and Huang, L. (1994) DNA transfection mediated by cationic liposomes containing lipopolysine: characterization and mechanism of action. Biochim. Biophys. Acta 1189, 195–203.PubMedCrossRefGoogle Scholar
  45. Zhu, J., Zhang, L., Hanisch, U.K., Fegner, P.L., and R.Reszka. (1996) A continuous intracereberal gene delivery system for in vivo liposome-mediated gene therapy. Gene Therapy 3, 472–476.PubMedGoogle Scholar
  46. Zhu, N., Liggitt, D., Liu, Y., and Debs, R. (1993) Systemic gene expression after intravenous DNA delivery into adult mice. Science 261, 209–211.PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1998

Authors and Affiliations

  • Natasha J. Caplen
    • 1
  1. 1.Clinical Gene Therapy Branch, National Human Genome Research InstituteNational Institutes of HealthBethesdaUSA

Personalised recommendations