Improved stability and efficacy of chitosan/pDNA complexes for gene delivery

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 charg...

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Bibliographic Details
Published in:Biotechnology letters 2015-03, Vol.37 (3), p.557-565
Main Authors: Cifani, Noemi, Chronopoulou, Laura, Pompili, Barbara, Di Martino, Antonio, Bordi, Federico, Sennato, Simona, Di Domenico, Enea Gino, Palocci, Cleofe, Ascenzioni, Fiorentina
Format: Article
Language:English
Subjects:
Applied Microbiology
Biochemistry
Biomedical and Life Sciences
Biotechnology
cations
Cell Line
Chitosan
Chitosan - metabolism
Computational efficiency
Deoxyribonucleic acid
DNA
DNA - metabolism
Drug Stability
Effectiveness
electrolytes
Epithelial Cells - metabolism
Gene therapy
Gene Transfer Techniques
Genes
Humans
In vitro testing
Life Sciences
Microbiology
Nanoparticles
Nanostructure
Original Research Paper
Plasmids
Polyelectrolytes
Polymers
Reproducibility of Results
Stability
Temperature
Time Factors
transfection
Transfection - methods
Transformation, Genetic
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Summary: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.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-014-1727-7