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Protein Transfection of Mouse Lung
Published on: May 15, 2013
Gene transfer into the lung by nanoparticle dextran-spermine/plasmid DNA complexes
Syahril Abdullah1, Wai Yeng Wendy-Yeo, Hossein Hosseinkhani
1UPM-MAKNA Cancer Research Laboratory, Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia. syahril@medic.upm.edu.my
Journal of Biomedicine & Biotechnology
|July 10, 2010
Summary
Dextran-spermine (D-SPM) enhances gene expression with plasmid DNA (pDNA). Optimal conditions were identified for in vitro and in vivo gene delivery, focusing on polymer-DNA ratios and delivery methods.
Area of Science:
- Biotechnology
- Gene Therapy
- Polymer Science
Background:
- Cationic polymers are investigated for gene delivery applications.
- Dextran-spermine (D-SPM) has shown potential for mediating gene expression.
- Optimization is needed for efficient D-SPM/plasmid DNA (pDNA) complex formation and delivery.
Purpose of the Study:
- To determine optimal conditions for D-SPM/pDNA gene expression in cell lines.
- To evaluate D-SPM/pDNA gene expression in mouse lungs via instillation.
- To characterize D-SPM/pDNA complex properties and their impact on gene delivery.
Main Methods:
- In vitro studies assessing D-SPM protection of pDNA from nuclease degradation.
- Optimization of D-SPM to pDNA weight-mixing ratios for gene transfer efficiency.
- In vivo gene expression analysis in mouse lungs following intranasal instillation of D-SPM/pDNA complexes.
Main Results:
- D-SPM partially protects pDNA from nuclease degradation.
- Optimal gene transfer efficiency in vitro achieved at a 12:1 D-SPM:pDNA weight ratio.
- In vivo gene expression in mouse lungs is dependent on D-SPM:pDNA ratio, pDNA amount, and time post-delivery.
- Complex size increases with pDNA amount, remaining in the nanometer range.
Conclusions:
- Dextran-spermine is an effective non-viral vector for gene delivery.
- Optimal formulation parameters are crucial for maximizing gene expression efficiency.
- D-SPM/pDNA complexes show promise for pulmonary gene delivery applications.
