Structure and dynamics of lipoplex formation examined using two-photon fluorescence cross-correlation spectroscopy
Structure and dynamics of lipoplex formation examined using two-photon fluorescence cross-correlation spectroscopy
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DOI：
10.1021/bi036133p
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发表时间：
2004-06-15
期刊：
影响因子：
2.9
通讯作者：
Cramb, DT
中科院分区：
文献类型：
--
作者：
Merkle, D;Lees-Miller, SP;Cramb, DT
形成可转染的脂质复合物所需的条件已被广泛研究[Zuhorn，I.美国，和Hoekstra，D.（2002）J. Membr. 189，167-179]。然而，到目前为止，实验还没有解决的顺序的事件在溶液中的脂质复合物形成或稳定的单囊泡脂质复合物中的每个囊泡的DNA分子的最大数量。在这项研究中，我们采用了双光子激发荧光相关光谱（TPE-FCS）和双光子荧光互相关光谱（TPE-XCS）来研究荧光标记的DNA和阳离子囊泡的结构和动力学同时。大的聚集的脂质复合物的形成对DNA-阳离子脂质电荷比的依赖性进行了测定，因为是40 bp的双链DNA寡核苷酸能够结合到一个单一的囊泡的最大数量。
The conditions required to form transfectable lipoplexes have been extensively studied [Zuhorn, I. S., and Hoekstra, D. (2002) J. Membr. Biol. 189, 167-179]. However, to date, experiments have not addressed either the order of events of lipoplex formation in solution or the maximum number of DNA molecules per vesicle in stable single-vesicle lipoplexes. In this study, we have employed two-photon excitation fluorescence correlation spectroscopy (TPE-FCS) and two-photon fluorescence cross-correlation spectroscopy (TPE-XCS) to examine both fluorescenced-labeled DNA and cationic vesicle structure and dynamics simultaneously. The dependence of large aggregated lipoplex formation on DNA-to-cationic lipid charge ratio was determined, as was the maximum number of 40 bp double-stranded DNA oligonucleotides able to bind to a single vesicle.