Skip to main content

Preparation and Physical Characterization of DNA-Binding Cationic Liposomes

  • Protocol
  • First Online:
Liposomes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1522))

Abstract

DNA-binding cationic liposomes are routinely employed as one of the major non-viral transfecting agents delivering DNA and other genes inside the cells. Cationic liposomes when complexed with DNA form a strong positively charged 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 in cationic liposomes.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. 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  PubMed  PubMed Central  Google Scholar 

  2. 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

    PubMed  Google Scholar 

  3. 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 

  4. 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  PubMed  Google Scholar 

  5. 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  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  7. 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  PubMed  Google Scholar 

  8. 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  PubMed  Google Scholar 

  9. 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  PubMed  Google Scholar 

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

    Article  Google Scholar 

  11. Pick U (1981) Liposomes with a large trapping capacity prepared by freezing and thawing of sonicated phospholipid mixtures. Arch Biochem Biophys 212(1):186–194

    Article  CAS  PubMed  Google Scholar 

  12. 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 

  13. 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 

  14. 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. Chem Phys Lipids 55(2):73–83

    Article  CAS  PubMed  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

© 2017 Springer Science+Business Media New York

About this protocol

Cite this protocol

Saxena, V. (2017). Preparation and Physical Characterization of DNA-Binding Cationic Liposomes. In: D'Souza, G. (eds) Liposomes. Methods in Molecular Biology, vol 1522. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6591-5_19

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-6591-5_19

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6589-2

  • Online ISBN: 978-1-4939-6591-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics