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Published on: March 1, 2013
Novel PEI/Poly-γ-Gutamic Acid Nanoparticles for High Efficient siRNA and Plasmid DNA Co-Delivery
Shu-Fen Peng1,2, Hung-Kun Hsu3, Chun-Cheng Lin4
1Department of Biological Science and Technology, China Medical University, Taichung 40402, Taiwan. sfpeng@mail.cmu.edu.tw.
Abstract:
The efficient delivery of sufficient amounts of nucleic acids into target cells is critical for successful gene therapy and gene knockdown. The DNA/siRNA co-delivery system has been considered a promising approach for cancer therapy to simultaneously express and inhibit tumor suppressor genes and overexpressed oncogenes, respectively, triggering synergistic anti-cancer effects. Polyethylenimine (PEI) has been identified as an efficient non-viral vector for transgene expression. In this study, we created a very high efficient DNA/siRNA co-delivery system by incorporating a negatively-charged poly-γ-glutamic acid (γ-PGA) into PEI/nucleic acid complexes. Spherical nanoparticles with about 200 nm diameter were formed by mixing PEI/plasmid DNA/siRNA/γ-PGA (dual delivery nanoparticles; DDNPs) with specific ratio (N/P/C ratio) and the particles present positive surface charge under all manufacturing conditions. The gel retardation assay shows both nucleic acids were effectively condensed by PEI, even at low N/P ratios. The PEI-based DDNPs reveal excellent DNA/siRNA transfection efficiency in the human hepatoma cell line (Hep 3B) by simultaneously providing high transgene expression efficiency and high siRNA silencing effect. The results indicated that DDNP can be an effective tool for gene therapy against hepatoma.
Insights
This study developed dual delivery nanoparticles (DDNPs) for efficient DNA and siRNA co-delivery in cancer therapy. These nanoparticles show high transfection efficiency for gene therapy and gene knockdown in hepatoma cells.
Area of Science:
- Biomedical Engineering
- Nanotechnology
- Molecular Biology
Background:
- Efficient nucleic acid delivery is crucial for gene therapy and gene knockdown.
- DNA/siRNA co-delivery offers synergistic anti-cancer effects by modulating tumor suppressor genes and oncogenes.
- Polyethylenimine (PEI) is an effective non-viral vector for transgene expression.
Purpose of the Study:
- To develop a highly efficient DNA/siRNA co-delivery system using PEI and poly-γ-glutamic acid (γ-PGA).
- To create dual delivery nanoparticles (DDNPs) for simultaneous gene expression and silencing.
- To evaluate the efficacy of DDNPs in hepatoma cells for cancer therapy.
Main Methods:
- Incorporation of negatively-charged γ-PGA into PEI/nucleic acid complexes to form DDNPs.
- Characterization of DDNPs (spherical nanoparticles, ~200 nm diameter, positive surface charge).
- Assessment of nucleic acid condensation using gel retardation assays and transfection efficiency in Hep 3B cells.
Main Results:
- PEI effectively condensed both DNA and siRNA, even at low N/P ratios.
- PEI-based DDNPs demonstrated excellent DNA/siRNA co-transfection efficiency in Hep 3B cells.
- Simultaneous high transgene expression and siRNA silencing effects were achieved.
Conclusions:
- DDNPs represent a highly efficient system for simultaneous DNA and siRNA delivery.
- The developed DDNPs show significant potential as a therapeutic tool for gene therapy against hepatoma.
- This co-delivery system offers a promising strategy for synergistic anti-cancer effects.
