Novel PEI/Poly-γ-Gutamic Acid Nanoparticles for High Efficient siRNA and Plasmid DNA Co-Delivery

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.

Development and In Vitro Evaluation of Linear PEI-Shelled Heparin/Berberine Nanoparticles in Human Osteosarcoma U-2 OS Cells

“…All experiments were performed in triplicate. The morphological changes of U-2 OS cells treated with BBR, HP/BBR, and HP/BBR/LPEI NPs were examined under a phase-contrast microscope [35].…”

“…Then, cells were washed with PBS twice, trypsinized, and transferred to FACS tubes. Subsequently, the uptake amount of BBR by U-2 OS cells was analyzed using a flow cytometer [35].…”

Systemic Delivery of Small Interfering RNA Therapeutics: Obstacles and Advances

“…Bioreducible polymer based targeted delivery of siRNA have also been revealed for human cancer gene therapy (Xia and Lin, 2012;Kozielski et al, 2014). Recent reports have shown co-delivery of siRNA and plasmid DNA using novel PEI/ poly--glutamic acid nanoparticles (Li et al, 2016;Peng et al, 2017). PLGA-PLL biodegradable nanoparticles exhibited controlled release, efficient gene silencing, endosomal escape, siRNA potency, and resistance against nuclease degradation (Zhou et al, 2012).…”

Preparation, characterization and evaluation of solid lipid nanoparticles and niosomes for ING4 gene delivery to MCF-7 cells

“…Complexation between cationic formulations and negative charged Plasmid DNA occurred via electrostatic interactions. Complexation efficiency of formulations were evaluated with gel retardation assay [23,24]. Different volumes of formulations and DNA solution were mixed and vortexed at 25°C for 30 minutes.…”