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

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

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.

• Language: English
• Keywords: gene electrotransfer, nanosecond pulses, high frequency bipolar pulses, electroporation, plasmid DNA
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: FE - Faculty of Electrical Engineering
• Publication status: Published
• Publication version: Version of Record
• Year: 2022
• Number of pages: 50 str.
• Numbering: Vol. 12, iss. 16, art. 8237
• PID: 20.500.12556/RUL-148017
• DOI: 10.3390/app12168237
• COBISS.SI-ID: 120304643
• Publication date in RUL: 20.07.2023

Record is a part of a journal

• Title: Applied sciences
• Shortened title: Appl. sci.
• Publisher: MDPI
• ISSN: 2076-3417
• COBISS.SI-ID: 522979353

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: genski elektroprenos, nanosekundni pulzi, visokofrekvenčni bipolarni pulzi, elektroporacija, plazmidna DNA