Influence of DNA condensation state on transfection efficiency in DNA/polymer complexes: An AFM and DLS comparative study

被引:43
|
作者
Volcke, C. [1 ]
Pirotton, S.
Grandfils, Ch.
Humbert, C.
Thiry, P. A.
Ydens, I.
Dubois, P.
Raes, M.
机构
[1] Univ Namur, LASMOS, B-5000 Namur, Belgium
[2] Univ Namur, Res Unit Cell Biol, B-5000 Namur, Belgium
[3] Univ Mons, LPCM, Mons, Belgium
[4] Univ Liege, Interfac Ctr Biomat, Liege, Belgium
关键词
atomic force microscopy; DNA; gene therapy vehicle; Dynamic Light Scattering;
D O I
10.1016/j.jbiotec.2006.02.010
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Atomic force microscopy (AFM) is used to describe the formation process of polymer/DNA complexes. Two main objectives of this research are presented. The first one is to apply AFM as an effective tool to analyse DNA molecules and different polycation/DNA complexes in order to evaluate their degree of condensation (size and shape). The other one is to search for a relationship between the condensation state of DNA and its transfection efficiency. In this study, linear methacrylate based polymers and globular SuperFect polymers are used in order to induce DNA condensation. Ternary complexes, composed of methacrylate based polymers and polyethylene glycol (PEG)-based copolymers, are also investigated. AFM allows us to confirm good condensation conditions and relate them (or not) to transfection efficiencies. These AFM results (obtained after drying in air) are compared with measurements deduced from Dynamic Light Scattering (DLS) experiments performed in water. This comparison allowed us to identify the structural modifications resulting from deposition on the mica surface. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:11 / 21
页数:11
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