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Fluorescence Resonance Energy Transfer (FRET)-Based Analysis of Lipoplexes

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Book cover Liposomes

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

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Abstract

We describe three applications of FRET technique to analyze structural and thermodynamic properties of cationic lipids and their complexes with DNA: (1) Lipid mixing assay to determine the degree to which individual vesicles undergo fusion upon complex formation. (2) DNA binding assay to obtain cationic lipid–DNA binding curves for thermodynamic analysis or binding stoichiometry characterization. (3) DNA spacing assay to determine changes in DNA packing in the multilamellar lipoplex particle.

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References

  1. Lakowicz JR (2004) Energy transfer. In: Principles of fluorescence spectroscopy, 3rd edn. Springer, New York, NY, pp 443–476

    Google Scholar 

  2. Kennedy MT, Pozharski EV, Rakhmanova VA, MacDonald RC (2000) Factors governing the assembly of cationic phospholipid-DNA complexes. Biophys J 78:1620–1633

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Pozharski E, MacDonald RC (2003) Lipoplex thermodynamics: determination of DNA-cationic lipoid interaction energies. Biophys J 85:3969–3978

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Madeira C, Loura LMS, Prieto M, Fedorov A, Aires-Barros MR (2008) Effect of ionic strength and presence of serum on lipoplexes structure monitorized by FRET. BMC Biotechnol 8:20

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Bartlett GR (1959) Phosphorus assay in column chromatography. J Biol Chem 234:466–468

    CAS  PubMed  Google Scholar 

  6. Duzgunes N, Allen TM, Fedor J, Papahadjopoulos D (1987) Lipid mixing during membrane aggregation and fusion: why fusion assays disagree. Biochemistry 26:8435–8442

    Article  CAS  PubMed  Google Scholar 

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Author information

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Center for Bimolecular Therapeutics, Institute for Bioscience and Biotechnology Research, University of Maryland School of Medicine, 9600 Gudelsky Dr., 4127D Bldg 2, Rockville, MD, 20850, USA

    Edwin Pozharski

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  1. Edwin Pozharski
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Corresponding author

Correspondence to Edwin Pozharski .

Editor information

Editors and Affiliations

  1. School of Pharmacy – Boston, MCPHS University, Boston, Massachusetts, USA

    Gerard G.M. D'Souza

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Pozharski, E. (2017). Fluorescence Resonance Energy Transfer (FRET)-Based Analysis of Lipoplexes. 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_20

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  • DOI: https://doi.org/10.1007/978-1-4939-6591-5_20

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  • 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

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