Abstract
Cationic liposomes are routinely employed as one of the major nonviral transfecting agents for intracellular delivery of hydrophilic molecules such as nucleic acids, peptides, and proteins. Cationic liposomes when complexed with DNA form a strong positively charged cationic liposome–DNA complex or lipoplex. The chapter discusses, primarily, the major preparation technique for cationic liposomes and its physical characterization, with a focus on SYBR Green I dye exclusion assay and DNA encapsulation enhancement by freeze–thaw technique. SYBR Green I dye exclusion assay is a technique to determine the total amount of liposomal lipids required to bind a unit weight of DNA, which is critical for transfection experiments. Freeze–thaw technique on the other hand is one of the major techniques to improve DNA encapsulation efficiency in liposomes.
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References
Lonez C, Vandenbranden M, Ruysschaert JM (2008) Cationic liposomal lipids: from gene carriers to cell signaling. Prog Lipid Res 47:340–347
Martin B, Sainlos M, Aissaoui A, Oudrhiri N, Hauchecorne M, Vigneron JP et al (2005) The design of cationic lipids for gene delivery. Curr Pharm Des 11:375–394
Felgner PL, Gadek TR, Holm M, Roman R, Chan HW, Wenz M, Northrop JP, Ringold GM, Danielsen M (1987) Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci U S A 84:7413–7417
Simões S, Pires P, Düzgünes N, Pedroso de Lima MC (1999) Cationic liposomes as gene transfer vectors: barriers to successful application in gene therapy. Curr Opin Mol Ther 1:147–157
Lasic DD, Strey HH, Stuart MCA, Podgornik R, Frederik PM (1997) The structure of DNA–liposome complexes. J Am Chem Soc 119:832–833
Radler JO, Koltover I, Salditt T, Safinya CR (1997) Structure of DNA–cationic liposome complexes. DNA intercalation in multi-lamellar membranes in distinct interhelical packing regimes. Science 275:810–814
Simberg D, Weisman S, Talmon Y, Barenholz Y (2004) DOTAP (and other cationic lipids): chemistry, biophysics, and transfection. Crit Rev Ther Drug Carrier Syst 21(4):257–317
Ewert K, Evans HM, Ahmad A, Slack NL, Lin AJ, Martin-Herranz A et al (2005) Lipoplex structures and their distinct cellular pathways. Adv Genet 53:119–155
Zuhorn IS, Engberts JB, Hoekstra D (2007) Gene delivery by cationic lipid vectors: overcoming cellular barriers. Eur Biophys J 36:349–362
Elouahabi A, Ruysschaert JM (2005) Formation and intracellular trafficking of lipoplexes and polyplexes. Mol Ther 11:336–347
Xu Y, Szoka FC (1996) Mechanism of DNA release from cationic liposome/DNA complexes used in cell transfection. Biochemistry 35:5616–5623
Weissig V, Lizano C, Torchilin VP (2000) Selective DNA release from DQAsome / DNA complexes at mitochondria-like membranes. Drug Deliv 7:1–5
Briggs C, Jones M (2005) SYBR green I-induced fluorescence in cultured immune cells: a comparison with Acridine Orange. Acta Histochem 107:301–312
Weissig V, D’Souza GGM, Torchilin VP (2001) DQAsomes/ DNA complexes release DNA upon contact with isolated mouse liver mitochondria. J Control Release 75:401–408
Wagle M, Martinville LE, D’Souza GGM (2011) The utility of an isolated mitochondrial fraction in the preparation of liposomes for the specific delivery of bioactives to mitochondria in live mammalian cells. Pharm Res 28:2790–2796
Pierre-Alain M, Oberholzer T, Luisi P (1997) Entrapment of nucleic acids in liposomes. Biochim Biophys Acta 1329:39–50
Pick U (1981) Liposomes with a large trap** capacity prepared by freezing and thawing of sonicated phospholipid mixtures. Arch Biochem Biophys 212(1):186–194
Schoen P, Bijl L, Wilschut J (1998) Efficient encapsulation of plasmid DNA in anionic liposomes by a freeze/thaw extrusion procedure. J Liposome Res 8(4):485–497
Zhou X, Klibanov AL, Huang L (1992) Improved encapsulation of DNA in pH-sensitive liposomes for transfection. J Liposome Res 2(1):125–139
Chapman CJ, Erdahl WL, Taylor RW, Pfeiffer DR (1990) Factors affecting solute entrapment in phospholipid vesicles prepared by the freeze-thaw extrusion method: a possible general method for improving the efficiency of entrapment. Chemistry of Physics and Lipids 55(2):73–83
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Saxena, V. (2023). Preparation and Physical Characterization of DNA Binding Cationic Liposomes. In: D'Souza, G.G., Zhang, H. (eds) Liposomes. Methods in Molecular Biology, vol 2622. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2954-3_10
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DOI: https://doi.org/10.1007/978-1-0716-2954-3_10
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