Preparation and Physical Characterization of DNA Binding Cationic Liposomes

  • Protocol
  • First Online:
Liposomes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2622))

  • 1444 Accesses

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.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
EUR 44.95
Price includes VAT (Germany)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
EUR 128.39
Price includes VAT (Germany)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
EUR 165.84
Price includes VAT (Germany)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free ship** worldwide - see info
Hardcover Book
EUR 235.39
Price includes VAT (Germany)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free ship** worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Lonez C, Vandenbranden M, Ruysschaert JM (2008) Cationic liposomal lipids: from gene carriers to cell signaling. Prog Lipid Res 47:340–347

    Article  CAS  Google Scholar 

  2. 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

    Article  CAS  Google Scholar 

  3. 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

    Article  CAS  Google Scholar 

  4. 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

    Google Scholar 

  5. Lasic DD, Strey HH, Stuart MCA, Podgornik R, Frederik PM (1997) The structure of DNA–liposome complexes. J Am Chem Soc 119:832–833

    Article  CAS  Google Scholar 

  6. 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

    Article  CAS  Google Scholar 

  7. 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

    Article  CAS  Google Scholar 

  8. 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

    Article  CAS  Google Scholar 

  9. Zuhorn IS, Engberts JB, Hoekstra D (2007) Gene delivery by cationic lipid vectors: overcoming cellular barriers. Eur Biophys J 36:349–362

    Article  CAS  Google Scholar 

  10. Elouahabi A, Ruysschaert JM (2005) Formation and intracellular trafficking of lipoplexes and polyplexes. Mol Ther 11:336–347

    Article  CAS  Google Scholar 

  11. Xu Y, Szoka FC (1996) Mechanism of DNA release from cationic liposome/DNA complexes used in cell transfection. Biochemistry 35:5616–5623

    Article  CAS  Google Scholar 

  12. Weissig V, Lizano C, Torchilin VP (2000) Selective DNA release from DQAsome / DNA complexes at mitochondria-like membranes. Drug Deliv 7:1–5

    Article  CAS  Google Scholar 

  13. Briggs C, Jones M (2005) SYBR green I-induced fluorescence in cultured immune cells: a comparison with Acridine Orange. Acta Histochem 107:301–312

    Article  Google Scholar 

  14. 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

    Article  CAS  Google Scholar 

  15. 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

    Article  CAS  Google Scholar 

  16. Pierre-Alain M, Oberholzer T, Luisi P (1997) Entrapment of nucleic acids in liposomes. Biochim Biophys Acta 1329:39–50

    Article  Google Scholar 

  17. 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

    Article  CAS  Google Scholar 

  18. 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

    Article  CAS  Google Scholar 

  19. Zhou X, Klibanov AL, Huang L (1992) Improved encapsulation of DNA in pH-sensitive liposomes for transfection. J Liposome Res 2(1):125–139

    Article  Google Scholar 

  20. 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

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Vaibhav Saxena .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

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

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-2954-3_10

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2953-6

  • Online ISBN: 978-1-0716-2954-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Navigation