Efficient gene transfection by electroporation - in vitro and in silico study of pulse parameters

Tjaša Potočnik (Author), Shaurya Sachdev (Author), Tamara Polajžer (Author), Alenka Maček Lebar (Author), Damijan Miklavčič (Author)

Abstract

Gene electrotransfer (GET) is a widely used method for nucleic acids’ delivery into cells. We explored, evaluated, and demonstrated the potential use of different pulse durations for introducing plasmid DNA (pDNA) into cells in vitro and compared the efficiency and dynamics of transgene expression after GET. We performed experiments on cell suspensions of 1306 fibroblasts and C2C12 myoblasts with four ranges of pulse durations (nanosecond, high frequency bipolar (HF-BP), and micro- and millisecond). Six different concentrations of pDNA encoding green fluorescent protein were used. We show that GET can be achieved with nanosecond pulses with a low pulse repetition rate (10 Hz). The GET’s efficiency depends on the pDNA concentration and cell line. Time dynamics of transgene expression are comparable between millisecond, microsecond, HF-BP, and nanosecond pulses but depend greatly on cell line. Lastly, based on the data obtained in the experiments of pDNA concentration effect on GET the model of the probability of pDNA and cell membrane contact during GET was developed. The model shows that pDNA migration is dominated by diffusion for nanosecond and HF-BP pulses and by electrophoresis for micro- and millisecond pulses. Modeling results can provide valuable guidance for further experiments and interpretations of the results obtained by various pulse protocols.

Keywords

genski elektroprenos;nanosekundni pulzi;visokofrekvenčni bipolarni pulzi;elektroporacija;plazmidna DNA;gene electrotransfer;nanosecond pulses;high frequency bipolar pulses;electroporation;plasmid DNA;

Data

Language:	English
Year of publishing:	2022
Typology:	1.01 - Original Scientific Article
Organization:	UL FE - Faculty of Electrical Engineering
UDC:	602.621
COBISS:	120304643
ISSN:	2076-3417
Views:	55
Downloads:	22
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	Slovenian
Secondary keywords:	genski elektroprenos;nanosekundni pulzi;visokofrekvenčni bipolarni pulzi;elektroporacija;plazmidna DNA;
Type (COBISS):	Article
Pages:	str. 1-51
Volume:	ǂiss. ǂ16
Issue:	8237
Chronology:	Aug.-2 2022
DOI:	10.3390/app12168237
ID:	19638981