title: Fluorescence Resonance Energy Transfer (FRET)-Based Analysis of Lipoplexes.
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.
authors.affiliation: 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. EPozharskiy@som.umaryland.edu.
authors.forename: Edwin
authors.initials: E
authors.lastname: Pozharski
mesh_terms: D001681:Biological Assay; D004247:DNA / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D008055:Lipids / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry*
chemical_list: D008055:Lipids; D008081:Liposomes; D004247:DNA
keywords: Binding assay; Cationic lipid; DNA spacing; FRET; Lipid mixing; Lipoplex; Vesicle fusion