Polymeric-DNA complexes (polyplexes) are constructed with multiple layers of counter-polyions as DNA/polyethylenimine/poly(acrylic acid)/polyethylenimine. The increased association of polyethylenimine achieved by the multilayer approach leads to substantial increases in expression of transgene for reporter plasmids without the need for excess free polymer typically required for non-viral gene delivery. This method of polyplex preparation provides the opportunity to improve transgene expression for gene therapy approaches to disease treatment.
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