Among polymeric polycations, chitosan has emerged as a powerful carrier for gene delivery. Only a few studies have focused on the stability of the chitosan/DNA complex under storage, although this is imperative for nanomedicinal applications. Here, we synthesized polyelectrolyte complexes at a charge ratio of 10 using 50 kDa chitosan and plasmid (p)DNA that encodes a GFP reporter. These preparations were stable up to 3 months at 4 °C and showed reproducible transfection efficiencies in vitro in HEK293 cells. In addition, we developed a methodology that increases the in vitro transfection efficiency of chitosan/pDNA complexes by 150 % with respect to standard procedures. Notably, intracellular pDNA release and transfected cells peaked 5 days following transection of mitotically active cells. These new developments in formulation technology enhance the potential for polymeric nanoparticle-mediated gene therapy.
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Istituto Pasteur Fondazione Cenci Bolognetti is acknowledged for financial support to FA and a Fellowship to EGD. LC, CP, SS and FB acknowledge financial support from Istituto Italiano di Tecnologia (IIT) Center of Life NanoScience-Sapienza. Conceived and designed the experiments: FA, CP, FB. Performed the experiments: NC, LC, BP, AM, SS. Analyzed the data: NC, LC, SS, EGD, FA, CP, FB. Wrote the paper: FA, CP.
Noemi Cifani, Laura Chronopoulou, and Barbara Pompili have contributed equally to the study.
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Cifani, N., Chronopoulou, L., Pompili, B. et al. Improved stability and efficacy of chitosan/pDNA complexes for gene delivery. Biotechnol Lett 37, 557–565 (2015). https://doi.org/10.1007/s10529-014-1727-7
- Chitosan/pDNA nanocomplexes
- DNA transfection
- Gene delivery
- HEK293 cells
- Transfection efficiency
- Therapeutic genes