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Evaluation of Polymeric Gene Delivery Nanoparticles by Nanoparticle Tracking Analysis and High-throughput Flow Cytometry
Published on: March 1, 2013
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A PEG-b-poly(disulfide-l-lysine) based redox-responsive cationic polymer for efficient gene transfection
Ihsan Ullah1, Jing Zhao, Shah Rukh
1School of Chemical Engineering and Technology, Tianjin University, Yaguan Road 135, Tianjin 300350, China. yakaifeng@tju.edu.cn.
Journal of Materials Chemistry. B
|April 8, 2020
Summary
Researchers developed a new bioreducible polymer, PEG-SSL, for gene therapy. This polymer efficiently delivers genes into cells with reduced toxicity, showing promise as a non-viral gene vector.
Area of Science:
- Biomaterials Science
- Gene Therapy
- Polymer Chemistry
Background:
- Gene therapy faces challenges with safety and low transfection efficiency of viral vectors.
- Redox-responsive bioreducible polymers offer potential as non-viral gene vectors due to controlled release and reduced toxicity.
Purpose of the Study:
- To synthesize and characterize a novel bioreducible poly(ethyleneglycol)-b-poly(disulfide-l-lysine) cationic polymer (PEG-SSL).
- To evaluate PEG-SSL as a non-viral gene vector for delivering plasmid ZNF580 (pZNF580) into cells.
Main Methods:
- PEG-SSL was synthesized via Michael addition reaction.
- PEG-SSL/pZNF580 polyplexes were formed and characterized for size, zeta potential, and stability against DNase I.
- In vitro transfection efficiency and cytotoxicity were assessed in EA.hy926 cells using flow cytometry and confocal microscopy.
Main Results:
- PEG-SSL formed nano-sized polyplexes with pZNF580, demonstrating efficient gene condensation and protection from degradation.
- The polymer showed rapid gene release in a reducing environment (5 mM DTT).
- PEG-SSL exhibited successful gene delivery and lower cytotoxicity compared to PEI25kDa in EA.hy926 cells, with good cellular uptake and nuclear co-localization.
Conclusions:
- PEG-SSL demonstrates potential as an effective and safe non-viral gene delivery vector.
- The bioreducible nature of PEG-SSL facilitates controlled gene release and reduces cellular toxicity.
- Further development of PEG-SSL could advance gene therapy applications.
