Journal
BIOTECHNOLOGY LETTERS
Volume 37, Issue 3, Pages 557-565Publisher
SPRINGER
DOI: 10.1007/s10529-014-1727-7
Keywords
Chitosan; Chitosan/pDNA nanocomplexes; DNA transfection; Gene delivery; HEK293 cells; Transfection efficiency; Therapeutic genes
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Funding
- Istituto Pasteur Fondazione Cenci Bolognetti
- Istituto Italiano di Tecnologia (IIT) Center of Life NanoScience-Sapienza
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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 A degrees 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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