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

C. Volcke; C. Humbert; P.A. Thiry; S. Pirotton; Martine Raes; I. Ydens; P. Dubois; Ch. Grandfils

doi:10.1016/j.jbiotec.2006.02.010

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

C. Volcke, C. Humbert, P.A. Thiry, S. Pirotton, Martine Raes, I. Ydens, P. Dubois, Ch. Grandfils

Résultats de recherche: Contribution à un journal/une revue › Article › Revue par des pairs

Résumé

La microscopie à force atomique (AFM) est utilisée pour décrire le processus de formation de complexes ADN/polymère. Les deux principaux objectifs de cette recherche sont présentés ci-àprès. Le premier consiste à appliquer l'AFM à l'analyse de molécules d'ADN et de différents complexes polycations/ADN afin d'évaluer leur degré de condensation (taille et forme). Le second objectif est de déterminer une relation entre l'état de condensation de l'ADN et son efficacité de transfection. Dans cette analyse, des polymères à base de méthacrylate linéaire et de polymères globulaires de SuperFect sont utilisés pour induire la condensation de l'ADN. Des complexes ternaires, composé de polymère à base de méthacrylate et de copolymère à base de polyéthylène glycol (PEG), sont étudiées. L'AFM nous a permis de confirmer les bonnes conditions de condensation et de les relier (ou pas) aux efficacités de transfection. Ces résultats AFM (obtenus après sèchage à l'air) sont comparés à des mesures déduites d'expériences de diffusion dynamique de lumière (DLS), réalisées en conditions liquide. Cette comparaison nous a permis d'identifier les modifications structurelles issues du dépôt à la surface du mica.

langue originale	Anglais
Pages (de - à)	11-21
Nombre de pages	11
journal	Journal of Biotechnology
Volume	125
Numéro de publication	1
Les DOIs	https://doi.org/10.1016/j.jbiotec.2006.02.010
Etat de la publication	Publié - 20 août 2006

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10.1016/j.jbiotec.2006.02.010

http://dx.doi.org/10.1016/j.jbiotec.2006.02.010

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

abstract = "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.",

keywords = " Dynamic light scattering, DNA, Gene therapy vehicle, Atomic force microscopy",

author = "C. Volcke and C. Humbert and P.A. Thiry and S. Pirotton and Martine Raes and I. Ydens and P. Dubois and Ch. Grandfils",

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T1 - Influence of DNA condensation state on transfection efficiency in DNA/polymer complexes

T2 - An AFM and DLS comparative study

AU - Volcke, C.

AU - Humbert, C.

AU - Thiry, P.A.

AU - Pirotton, S.

AU - Raes, Martine

AU - Ydens, I.

AU - Dubois, P.

AU - Grandfils, Ch.

N1 - MEDLINE® is the source for the MeSH terms of this document.

PY - 2006/8/20

Y1 - 2006/8/20

N2 - 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.

AB - 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.

KW - Dynamic light scattering

KW - DNA

KW - Gene therapy vehicle

KW - Atomic force microscopy

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DO - 10.1016/j.jbiotec.2006.02.010

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EP - 21

JO - Journal of Biotechnology

JF - Journal of Biotechnology

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

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