Fluorescence resonance energy transfer: Evaluation of the intracellular stability of polyplexes
Fluorescence resonance energy transfer: Evaluation of the intracellular stability of polyplexes
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DOI：
10.1016/j.ejpb.2006.01.012
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发表时间：
2006-06-01
影响因子：
4.9
通讯作者：
Goepferich, Achim
中科院分区：
文献类型：
--
作者：
Breunig, Miriam;Lungwitz, Uta;Goepferich, Achim
对非病毒核酸递送系统细胞内机制的研究为其有效性的提高提供了巨大的动力。尤其是非病毒载体系统的细胞内核酸释放可能是某些聚合物高转染率的相关标准。因此，我们评价了荧光共振能量转移(FRET)结合激光共聚焦扫描显微镜或流式细胞术作为工具来确定由质粒DNA和线性聚乙烯亚胺构建的多聚体的细胞内崩解。在显微镜中，可以观察单个细胞内的多个复合体，敏化发射测量和受体光漂白已经被测试用于定量FRET分析。相反，用基于流式细胞术的方法分析整个细胞群体。我们认为，FRET是一种有用的工具来评估不同聚合物构建的复合体的细胞内崩解。(C)2006爱思唯尔B.V.保留所有权利。
The investigation of intracellular mechanisms of non-viral nucleic acid delivery systems has provided great impetus for the improvement of their efficacy. Especially the intracellular release of the nucleic acid from the non-viral carrier system may be a relevant criterion for the high transfection efficiency of certain polymers. Therefore, we evaluated fluorescence resonance energy transfer (FRET) in combination with confocal laser scanning microscopy or flow cytometry as tool to determine the intracellular disintegration of polyplexes built with plasmid DNA and linear polyethylenimine. In microscopy, which allowed for an observation of polyplexes within single cells, sensitized emission measurement and acceptor photobleaching have been tested towards quantitative FRET analysis. In contrast, the whole cell population was analyzed by the flow cytometry-based method. We suggest that FRET is a useful tool to evaluate the intracellular disintegration of polyplexes built with various polymers. (c) 2006 Elsevier B.V. All rights reserved.