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基于荧光共振能量转移（FRET）的脂质体分析

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

作者信息

Pozharski Edwin

机构信息

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.

出版信息

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

DOI:10.1007/978-1-4939-6591-5_20

PMID:27837545

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.

摘要

我们描述了荧光共振能量转移（FRET）技术在分析阳离子脂质及其与DNA复合物的结构和热力学性质方面的三种应用：（1）脂质混合测定，以确定单个囊泡在复合物形成时发生融合的程度。（2）DNA结合测定，以获得用于热力学分析或结合化学计量表征的阳离子脂质-DNA结合曲线。（3）DNA间距测定，以确定多层脂质体颗粒中DNA堆积的变化。