Abstract
Entrapment of plasmid DNA (pDNA) in an aqueous compartment separated from the bulk external aqueous medium by a phospholipid bilayer resembles a structure similar to a primitive living cell, and interestingly, this phenomenon occurs completely self-assembled. Being inspired by such a structure as well as using the dehydration–rehydration technique, we were able to encapsulate pDNA without using multivalent cations and with high efficiency (98 %) into noncationic lipid bilayer vesicles. These liposomes which were composed of dimyristoyl-sn-glycero-3-phosphocholine unlike cationic liposomes, were nontoxic. The obtained liposome structure was able protect DNA against nuclease and was completely stable, in a way that even after 6 months, it still kept the pDNA in its structure, and there was a small change in its size (100–150 nm) determined by dynamic light scattering. The purpose of this research is to polarize the researchers’ interest toward utilization of neutral liposomes originating from the cell membrane as the most efficient carrier for gene delivery. We indicated that in using such carriers, which are the most similar synthetic structures to viruses, their inability in electrostatic interaction with DNA would not be an obstacle for entrapping nucleic acids.
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References
Pisani, M., Mobbili, G., & Bruni, P. (2011). Neutral liposomes and DNA transfection. In X. Yuan (Ed.), Non-viral gene therapy (pp. 319–348). Shanghai: InTech.
Elsabahy, M., Nazarali, A., & Foldvari, M. (2011). Non-viral nucleic acid delivery: Key challenges and future directions. Current Drug Delivery, 8, 235–244.
Mintzer, M. A., & Simanek, E. E. (2009). Nonviral vectors for gene delivery. Chemical Reviews, 109, 259–302.
MacLachlan, I., Cullis, P. R., & Graham, R. W. (2000). Synthetic virus systems for systemic gene therapy. In N. Smythe-Templeton & D. Lasic (Eds.), Gene therapy: Therapeutic mechanisms and strategies (pp. 267–290). New York: Marcel Dekker.
Tresset, G., Cheong, W. C., Tan, Y. L., Boulaire, J., & Lam, Y. M. (2007). Phospholipid-based artificial viruses assembled by multivalent cations. Biophysical Journal, 93, 637–644.
Ulrich, A. S. (2002). Biophysical aspects of using liposomes as delivery vehicles. Bioscience Reports, 22, 129–150.
Wilschut, J., & Hoekstra, D. (1984). Membrane fusion: From liposomes to biological membranes. Trends in Biochemical Sciences, 9, 479–483.
Deamer, D. (2005). A giant step towards artificial life? Trends in Biotechnology, 23, 336–338.
Dzieciol, A. J., & Mann, S. (2012). Designs for life: Protocell models in the laboratory. Chemical Society Reviews, 41, 79–85.
Walde, P. (2010). Building artificial cells and protocell models: Experimental approaches with lipid vesicles. BioEssays, 32, 296–303.
Mayer, L. D., Bally, M. B., Hope, M. J., & Cullis, P. R. (1986). Techniques for encapsulating bioactive agents into liposomes. Chemistry and Physics of Lipids, 40, 333–345.
Monnard, P. A., Luptak, A., & Deamer, D. W. (2007). Models of primitive cellular life: Polymerases and templates in liposomes. Philosophical Transactions of the Royal Society of London. Series B, Biological sciences, 362, 1741–1750.
Mugabe, C., Azghani, A., & Omri, A. (2006). Preparation and characterization of dehydration–rehydration vesicles loaded with aminoglycoside and macrolide antibiotics. International Journal of Pharmaceutics, 307, 244–250.
Knoll, G., Burger, K. N., Bron, R., van Meer, G., & Verkleij, A. J. (1988). Fusion of liposomes with the plasma membrane of epithelial cells: Fate of incorporated lipids as followed by freeze fracture and autoradiography of plastic sections. Journal of Cell Biology, 107, 2511–2521.
Sternberg, B., Gumpert, J., Meyer, H. W., & Reinhardt, G. (1986). Structures of liposome membranes as models for similar features of cytoplasmic membranes of bacteria. Acta Histochemica Supplement, 33, 139–145.
Valenzuela, S. M. (2007). Liposome techniques for synthesis of biomimetic lipid membranes. In D. Martin (Ed.), Nanobiotechnology of biomimetic membranes (pp. 75–87). New York: Springer.
Wiethoff, C. M., Gill, M. L., Koe, G. S., Koe, J. G., & Russell, M. C. (2002). The structural organization of cationic lipid–DNA complexes. Journal of Biological Chemistry, 277, 44980–44987.
Fraley, R., Subramani, S., Berg, P., & Papahadjopoulos, D. (1980). Introduction of liposome-encapsulated SV40 DNA into cells. Journal of Biological Chemistry, 255, 10431–10435.
Kudsiova, L., Arafiena, C., & Lawrence, M. J. (2008). Characterisation of chitosan-coated vesicles encapsulating DNA suitable for gene delivery. Journal of Pharmaceutical Sciences, 97, 3981–3997.
Gregoriadis, G., Saffie, R., & Hart, S. L. (1996). High yield incorporation of plasmid DNA within liposomes: Effect on DNA integrity and transfection efficiency. Journal of Drug Targeting, 3, 469–475.
Gregoriadis, G., Bacon, A., Caparros-Wanderley, W., & McCormack, B. (2003). Plasmid DNA vaccines: Entrapment into liposomes by dehydration–rehydration. Methods in Enzymology, 367, 70–80.
Maurer, S. E., & Monnard, P. A. (2011). Primitive membrane formation, characteristics and roles in the emergent properties of a protocell. Entropy, 13, 466–484.
Monnard, P. A., & Deamer, D. W. (2001). Nutrient uptake by protocells: A liposome model system. Origins of Life and Evolution of the Biosphere, 31, 145–155.
Bruni, P., Francescangeli, O., Marini, M., Mobbili, G., Pisani, M., & Smorlesi, A. (2011). Can neutral liposomes be considered as genetic material carriers for human gene therapy? Mini-Reviews in Organic Chemistry, 8, 38–48.
Mozafari, M. R., & Omri, A. (2007). Importance of divalent cations in nanolipoplex gene delivery. Journal of Pharmaceutical Sciences, 96, 1955–1966.
Pisani, M., Bruni, P., Caracciolo, G., Caminiti, R., & Francescangeli, O. (2006). Structure and phase behavior of self-assembled DPPC–DNA–metal cation complexes. Journal of Physical Chemistry B, 110, 13203–13211.
Suleymanoglu, E. (2006). Phospholipid–nucleic acid recognition: Energetics of DNA–Mg2+–phosphatidylcholine ternary complex formation and its further compaction as a gene delivery formulation. PDA Journal of Pharmaceutical Science and Technology, 60, 218–231.
Bruni, P., Pisani, M., Amici, A., Marchini, C., Montani, M., & Francescangeli, O. (2006). Self-assembled ternary complexes of neutral liposomes, deoxyribonucleic acid, and bivalent metal cations. Promising vectors for gene transfer? Applied Physics Letters, 88, 073901–073903.
Kuvichkin, V. V. (2009). Investigation of ternary complexes: DNA–phosphatidylcholine liposomes–Mg2+ by freeze-fracture method and their role in the formation of some cell structures. Journal of Membrane Biology, 231, 29–34.
Manosroi, A., Thathang, K., Werner, R. G., Schubert, R., & Manosroi, J. (2008). Stability of luciferase plasmid entrapped in cationic bilayer vesicles. International Journal of Pharmaceutics, 356, 291–299.
Perrie, Y., & Gregoriadis, G. (2000). Liposome-entrapped plasmid DNA: Characterisation studies. Biochimica et Biophysica Acta, 1475, 125–132.
Kurihara, K., Tamura, M., Shohda, K., Toyota, T., Suzuki, K., & Sugawara, T. (2011). Self-reproduction of supramolecular giant vesicles combined with the amplification of encapsulated DNA. Nature Chemistry, 3, 775–781.
Pupo, E., Padrón, A., Santana, E., Sotolongo, J., Quintana, D., Dueñas, S., et al. (2005). Preparation of plasmid DNA-containing liposomes using a high-pressure homogenization–extrusion technique. Journal of Controlled Release, 104, 379–396.
Gundermann, K., & Scheele, E. (2003). Polyunsaturated phosphatidylcholine in chronic liver disease? Past and present. In B. F. Szuhaj & W. van Nieuwenhuyzen (Eds.), Nutrition and biochemistry of phospholipids (pp. 152–162). Urbana, IL: AOCS Publishing.
Tandy, S., Chung, R. W. S., Kamili, A., Wat, E., Weir, J. M., Meikle, P. J., et al. (2010). Hydrogenated phosphatidylcholine supplementation reduces hepatic lipid levels in mice fed a high-fat diet. Atherosclerosis, 213, 142–147.
Schuck, S., Honsho, M., Ekroos, K., Shevchenko, A., & Simons, K. (2003). Resistance of cell membranes to different detergents. Proceedings of the National Academy of Sciences of the United States of America, 100, 5795–7800.
Sot, J., Collado, M. I., Arrondo, J. L. R., Alonso, A., & Goñi, F. M. (2002). Triton X-100-resistant bilayers: Effect of lipid composition and relevance to the raft phenomenon. Langmuir, 18, 2828–2835.
Koynova, R., & Tenchov, B. (2001). Interactions of surfactants and fatty acids with lipids. Current Opinion in Colloid & Interface Science, 6, 277–286.
London, E., & Brown, D. A. (2000). Insolubility of lipids in triton X-100: Physical origin and relationship to sphingolipid/cholesterol membrane domains (rafts). Biochimica et Biophysica Acta, 1508, 182–195.
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The authors would like to express their gratefulness to the Research Council of Tarbiat Modares University for providing financial support for this study.
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Ramezani, R., Sadeghizadeh, M., Behmanesh, M. et al. Characterization of Zwitterionic Phosphatidylcholine-Based Bilayer Vesicles as Efficient Self-Assembled Virus-Like Gene Carriers. Mol Biotechnol 55, 120–130 (2013). https://doi.org/10.1007/s12033-013-9663-7
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DOI: https://doi.org/10.1007/s12033-013-9663-7