Imaging in solution of (Lys)16-containing bifunctional synthetic peptide/DNA nanoparticles for gene delivery
Imaging in solution of (Lys)16-containing bifunctional synthetic peptide/DNA nanoparticles for gene delivery
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DOI：
10.1016/j.bbagen.2004.02.004
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发表时间：
2004-04-07
影响因子：
3
通讯作者：
Fabre, JW
中科院分区：
文献类型：
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作者：
Collins, L;Kaszuba, M;Fabre, JW
非病毒载体/DNA纳米颗粒在生理水溶液中的物理性质知之甚少。通常用于分析工业和环境流体的流体颗粒图像分析仪（FPIA）用于可视化含有单个（Lys）（16）的肽/DNA颗粒。使用含八个（Lys）（16）的合成肽以2.8：1的恒定+-电荷比与10 μ g/ml的质粒DNA在磷酸盐缓冲盐水中产生肽/DNA颗粒。还进行了动态光散射（DLS）和基因递送研究。我们提出了第一个图像的非病毒载体/DNA纳米粒子在生理水溶液中，连同精确测量的单个颗粒的大小和形状的解决方案，并为第一次，一个准确的测量颗粒的数量。不同肽的颗粒大小和形状、颗粒数量和基因递送效率显著不同。在体外基因传递的标准条件下，我们估计每个靶细胞大约有60个肽/DNA纳米颗粒，每个纳米颗粒含有大约70,000个质粒。这种在溶液中对单个载体/DNA纳米颗粒进行成像并准确计数的新能力将使非病毒基因递送系统的评估更加精确，并对基因递送实验进行更定量的解释。(C)2004 Elsevier B. V.保留所有权利。
The physical properties of non-viral vector/DNA nanoparticles in physiological aqueous solution are poorly understood. A Fluid Particle image Analyser (FPIA), normally used for analysis of industrial and environmental fluids, was used to visualise individual (Lys)(16)-containing peptide/DNA particles. Eight (Lys)(16)-containing synthetic peptides were used to generate peptide/DNA particles at a constant + to - charge ratio of 2.8:1 with 10 mug/ml of plasmid DNA in phosphate buffered saline. Dynamic Light Scattering (DLS) and gene delivery studies were also performed. We present the first images of non-viral vector/DNA nanoparticles in physiological aqueous solution, together with precise measurements of individual particle size and shape in solution and, for the first time, an accurate measure of particle number. Particle size and shape, particle number, and efficiency for gene delivery varied markedly with different peptides. Under standard conditions for in vitro gene delivery, we estimate similar to60 peptide/DNA nanoparticles per target cell, each containing similar to70,000 plasmids. This novel capacity to image individual vector/DNA nanoparticles in solution and to count them accurately will enable a more precise assessment of non-viral gene delivery systems, and a more quantitative interpretation of gene delivery experiments. (C) 2004 Elsevier B.V. All rights reserved.