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Polymers 2016, 8(6), 224; doi:10.3390/polym8060224

Influence of Polyplex Formation on the Performance of Star-Shaped Polycationic Transfection Agents for Mammalian Cells

1
Process Biotechnology, University of Bayreuth, 95440 Bayreuth, Germany
2
Simpson Querrey Institute for BioNanotechnology, Northwestern University, Chicago, 60611 IL, USA
3
Institute of Organic Chemistry, Johannes-Gutenberg-University, 55099 Mainz, Germany
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editor: Roland G. Winkler
Received: 12 April 2016 / Revised: 30 May 2016 / Accepted: 1 June 2016 / Published: 6 June 2016
(This article belongs to the Collection Polyelectrolytes)
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Abstract

Genetic modification (“transfection”) of mammalian cells using non-viral, synthetic agents such as polycations, is still a challenge. Polyplex formation between the DNA and the polycation is a decisive step in such experiments. Star-shaped polycations have been proposed as superior transfection agents, yet have never before been compared side-by-side, e.g., in view of structural effects. Herein four star-shaped polycationic structures, all based on (2-dimethylamino) ethyl methacrylate (DMAEMA) building blocks, were investigated for their potential to deliver DNA to adherent (CHO, L929, HEK-293) and non-adherent (Jurkat, primary human T lymphocytes) mammalian cells. The investigated vectors included three structures where the PDMAEMA arms (different arm length and grafting densities) had been grown from a center silsesquioxane or silica-coated γ-Fe2O3-core and one micellar structure self-assembled from poly(1,2-butadiene)-block PDMAEMA polymers. All nano-stars combined high transfection potential with excellent biocompatibility. The micelles slightly outperformed the covalently linked agents. For method development and optimization, the absolute amount of polycation added to the cells was more important than the N/P-ratio (ratio between polycation nitrogen and DNA phosphate), provided a lower limit was passed and enough polycation was present to overcompensate the negative charge of the plasmid DNA. Finally, the matrix (NaCl vs. HEPES-buffered glucose solution), but also the concentrations adjusted during polyplex formation, affected the results. View Full-Text
Keywords: gene delivery; mammalian cells; non-viral; PDMAEMA; T lymphocytes; transfection gene delivery; mammalian cells; non-viral; PDMAEMA; T lymphocytes; transfection
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Raup, A.; Stahlschmidt, U.; Jérôme, V.; Synatschke, C.V.; Müller, A.H.E.; Freitag, R. Influence of Polyplex Formation on the Performance of Star-Shaped Polycationic Transfection Agents for Mammalian Cells. Polymers 2016, 8, 224.

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