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Sustained expression in mammalian cells with DNA complexed with chitosan nanoparticles
Xiong Wei Li1, Daniel Ken Lem Lee, Albert Sun Chi Chan
1The School of Pharmacy, University of London, 29-39 Brunswick Square, London WC1N 1AX, UK.
Biochimica Et Biophysica Acta
|October 29, 2003
Summary
Chitosan nanoparticles show potential as effective gene delivery vectors, offering sustained gene expression in mammalian cells for up to 10 days without toxicity. These chitosan systems outperform commercial reagents in duration and safety.
Area of Science:
- Biomaterials Science
- Gene Therapy
- Nanotechnology
Background:
- Gene transfection is crucial for gene therapy and research.
- Developing safe and efficient non-viral gene delivery vectors is a key challenge.
- Chitosan, a natural polymer, offers potential for gene delivery due to its biocompatibility and biodegradability.
Purpose of the Study:
- To investigate the preparation and in vitro efficiency of chitosan-based nanoparticles for gene transfection.
- To evaluate the performance of chitosan nanoparticles as gene delivery vectors in mammalian cells.
- To compare the efficacy and cytotoxicity of chitosan vectors with commercial transfection reagents.
Main Methods:
- Chitosan nanoparticles were prepared and characterized for size using photon correlation spectroscopy (PCS) and transmission electron microscopy (TEM).
- DNA complexation ability was assessed using gel retardation assays with reporter plasmids (pGL3-Control, pCH110).
- Transfection efficiency was evaluated in 293 human kidney cells and CHO-K1 cells, measuring luciferase gene expression and comparing with SuperFect and Lipofectin.
Main Results:
- Chitosan nanoparticles ranged from 40-200 nm (PCS) and 40-80 nm (TEM).
- Chitosan-DNA complexes demonstrated sustained luciferase gene expression for approximately 10 days in transfected cells.
- Chitosan showed no significant cytotoxicity, unlike SuperFect and Lipofectin, which exhibited marked cytotoxicity.
Conclusions:
- Chitosan nanoparticles are effective and safe non-viral vectors for gene delivery into mammalian cells.
- The sustained gene expression achieved with chitosan surpasses that of commercial reagents.
- Chitosan's potential for gene therapy applications is highlighted by its efficiency, safety, and prolonged expression profile.