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

Methods Mol Biol. 2017;1522:251-256. doi: 10.1007/978-1-4939-6591-5_20.

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.

Keywords: Binding assay; Cationic lipid; DNA spacing; FRET; Lipid mixing; Lipoplex; Vesicle fusion.

MeSH terms

  • Biological Assay
  • DNA / metabolism
  • Fluorescence Resonance Energy Transfer / methods*
  • Lipids / chemistry
  • Liposomes / chemistry*

Substances

  • Lipids
  • Liposomes
  • DNA