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

Title data

Raup, Alexander ; Stahlschmidt, Ullrich ; Jérôme, Valérie ; Synatschke, Christopher V. ; Müller, Axel H. E. ; Freitag, Ruth:
Influence of Polyplex Formation on the Performance of Star-Shaped Polycationic Transfection Agents for Mammalian Cells.
Process Biotechnology, University of Bayreuth, 95440 Bayreuth, Germany; Simpson Querrey Institute for BioNanotechnology, Northwestern University, Chicago, 60611 IL, USA; Institute of Organic Chemistry, Johannes-Gutenberg-University, 55099 Mainz, Germany
In: Polymers. Vol. 8 (June 2016) Issue 6 . - p. 224.
ISSN 2073-4360
DOI: https://doi.org/10.3390/polym8060224

Official URL: Volltext

Abstract in another language

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.

Further data

Item Type: Article in a journal
Refereed: Yes
Keywords: gene delivery; mammalian cells; non-viral; PDMAEMA; T lymphocytes; transfection
Institutions of the University: Faculties
Faculties > Faculty of Biology, Chemistry and Earth Sciences
Faculties > Faculty of Engineering Science
Faculties > Faculty of Engineering Science > Chair Process Biotechnology
Faculties > Faculty of Engineering Science > Chair Process Biotechnology > Chair Process Biotechnology - Univ.-Prof. Dr. Ruth Freitag
Result of work at the UBT: Yes
DDC Subjects: 500 Science
500 Science > 500 Natural sciences
600 Technology, medicine, applied sciences
600 Technology, medicine, applied sciences > 610 Medicine and health
600 Technology, medicine, applied sciences > 620 Engineering
Date Deposited: 07 Jun 2016 08:28
Last Modified: 07 Jun 2016 08:28
URI: https://eref.uni-bayreuth.de/id/eprint/32539