The impact of impaired DNA mobility on gene electrotransfer efficiency: analysis in 3D model

BioMedical Engineering OnLine

Saša Haberl Meglič¹  Mojca Pavlin² 

[1] Faculty of Electrical Engineering, Laboratory of Biocybernetics, University of Ljubljana, Tržaška 25, 1000, Ljubljana, Slovenia;Faculty of Medicine, Institute of Biophysics, University of Ljubljana, Vrazov trg 2, 1000, Ljubljana, Slovenia;Faculty of Electrical Engineering, Group for Nano and Biotechnological Applications, University of Ljubljana, Tržaška 25, 1000, Ljubljana, Slovenia;

关键词: 3D in vitro model;    Collagen gel;    Diffusion;    Electrophoresis;    Gene electrotransfer;    GFP;    High-voltage pulse;    Low-voltage pulse;    Different polarity pulses;   

DOI  :  10.1186/s12938-021-00922-3

来源: Springer

【 摘 要 】

BackgroundGene electrotransfer is an established method that enables transfer of DNA into cells with electric pulses. Several studies analyzed and optimized different parameters of gene electrotransfer, however, one of main obstacles toward efficient electrotransfection in vivo is relatively poor DNA mobility in tissues. Our aim was to analyze the effect of impaired mobility on gene electrotransfer efficiency experimentally and theoretically. We applied electric pulses with different durations on plated cells, cells grown on collagen layer and cells embedded in collagen gel (3D model) and analyzed gene electrotransfer efficiency. In order to analyze the effect of impaired mobility on gene electrotransfer efficiency, we applied electric pulses with different durations on plated cells, cells grown on collagen layer and cells embedded in collagen gel (3D model) and analyzed gene electrotransfer efficiency.ResultsWe obtained the highest transfection in plated cells, while transfection efficiency of embedded cells in 3D model was lowest, similarly as in in vivo. To further analyze DNA diffusion in 3D model, we applied DNA on top or injected it into 3D model and showed, that for the former gene electrotransfer efficiency was similarly as in in vivo. The experimental results are explained with theoretical analysis of DNA diffusion and electromobility.ConclusionWe show, empirically and theoretically that DNA has impaired electromobility and especially diffusion in collagen environment, where the latter crucially limits electrotransfection. Our model enables optimization of gene electrotransfer in in vitro conditions.

【 授权许可】

CC BY