Promoter, transgene, and cell line effects in the transfection of mammalian cells using PDMAEMA-based nano-stars

Alexander Raup; Valérie Jérôme; Ruth Freitag; Christopher V Synatschke; Axel H E Müller

doi:10.1016/j.btre.2016.05.003

Promoter, transgene, and cell line effects in the transfection of mammalian cells using PDMAEMA-based nano-stars

Biotechnol Rep (Amst). 2016 Jun 16:11:53-61. doi: 10.1016/j.btre.2016.05.003. eCollection 2016 Sep.

Authors

Alexander Raup¹, Valérie Jérôme¹, Ruth Freitag¹, Christopher V Synatschke², Axel H E Müller²

Affiliations

¹ Process Biotechnology, University of Bayreuth, 95440 Bayreuth, Germany.
² Macromolecular Chemistry II, University of Bayreuth, 95440 Bayreuth, Germany.

Abstract

Non-viral transfection protocols are typically optimized using standard cells and reporter proteins, potentially underestimating cellular or transgene effects. Here such effects were studied for two human (Jurkat, HEK-293) and two rodent (CHO-K1, L929) cell lines and three fluorescent reporter proteins. Expression of the enhanced green fluorescent protein (EGFP) was studied under the control of the human elongation factor 1 alpha promoter and three viral promoters (SV40, SV40/enhancer, CMV), that of ZsYellow1 (yellow fluorescence) and mCherry (red fluorescence) for the CMV promoter. Results varied with the cell line, in particular for the Jurkat cells. Pair-wise co-transfection of the CMV controlled transgenes resulted in a significant fraction of monochromatic cells (EGFP for EGFP/YFP and EGFP/RFP co-transfections, YFP in case of YFP/RFP co-transfections). Only Jurkat cells were almost incapable of expressing YFP. Dilution of the plasmid DNA with a non-expressed plasmid showed cell line dependent effects on transfection efficiency and/or expression levels.

Keywords: Co-expression; Fluorescent reporter protein; Mammalian cell; Non-viral gene delivery; Poly(2-dimethylamino) ethyl methacrylate.