Chitosan-Based Vector/DNA Complexes for Gene Delivery: Biophysical Characteristics and Transfection Ability
- Patrick ErbacherAffiliated withLaboratoire de Chimie Génétique, Université Louis-Pasteur de Strasbourg
- , Shaomin ZouAffiliated withLaboratoire de Chimie Génétique, Université Louis-Pasteur de Strasbourg
- , Thierry BettingerAffiliated withLaboratoire de Chimie Génétique, Université Louis-Pasteur de Strasbourg
- , Anne-Marie SteffanAffiliated withInstitut de Virologie
- , Jean-Serge RemyAffiliated withLaboratoire de Chimie Génétique, Université Louis-Pasteur de Strasbourg
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Purpose. Chitosan, a natural cationic polysaccharide, is a candidate non-viral vector for gene delivery. With the aim of developing this system, various biophysical characteristics of chitosan-condensed DNA complexes were measured, and transfections were performed.
Methods. Transmission electronic microscopy (TEM) visualizations, sedimentation experiments, dynamic light scattering (DLS), and zeta potential measurements were realized. Transfections were made by using the luciferase reporter gene.
Results. In defined conditions, plasmid DNA formulated with chitosan produced homogenous populations of complexes which were stable and had a diameter of approximately 50−100 nm. Discrete particles of nicely condensed DNA had a donut, rod, or even pretzel shape. Chitosan/DNA complexes efficiently transfected HeLa cells, independently of the presence of 10% serum, and did not require an added endosomolytic agent. In addition, gene expression gradually increased over time, from 24 to 96 hours, whereas in the same conditions the efficacy of polyethylenimine-mediated transfection dropped by two orders of magnitude. At 96 hours, chitosan was found to be 10 times more efficient than PEI. However, chitosan-mediated transfection depended on the cell type. This dependency is discussed here.
Conclusions. Chitosan presents some characteristics favorable for gene delivery, such as the ability to condense DNA and form small discrete particles in defined conditions.
- Chitosan-Based Vector/DNA Complexes for Gene Delivery: Biophysical Characteristics and Transfection Ability
Volume 15, Issue 9 , pp 1332-1339
- Cover Date
- Print ISSN
- Online ISSN
- Kluwer Academic Publishers-Plenum Publishers
- Additional Links
- gene therapy
- gene transfer
- cationic polymer
- Industry Sectors
- Author Affiliations
- 1. Laboratoire de Chimie Génétique, Université Louis-Pasteur de Strasbourg, CNRS UMR 7514, Faculté de Pharmacie, BP24, F-67401, Illkirch, France
- 2. Institut de Virologie, INSERM U74, Strasbourg, France