Expression of GFP and DsRed fluorescent proteins after gene electrotransfer of tumour cells in vitro

Komel, Tilen (author), Bošnjak, Maša (author), Serša, Gregor (author), Čemažar, Maja (author)

Fluorescent reporter genes are widely used to study the transfection of various types of primary cells and cell lines. The aim of our research was to investigate the expression dynamics of GFP and DsRed reporter genes individually and combined after gene electrotransfer of plasmids with two different electroporation protocols in B16F10 and CT26 cells in vitro. The cytotoxicity after gene electrotransfer of both plasmids was first determined. Second, the intensity of fluorescence and the percentage of cells transfected with both plasmids individually and in combination were monitored in real time. The results show that the percentage of viability after gene electrotransfer of plasmids using the EP2 pulses was significantly higher compared to the EP1 pulses. In contrast, the percentage of transfected cells and fluorescence intensity were higher after gene electrotransfer with the EP1 pulse protocol. Moreover, the percentage of transfected cells was higher and started earlier in the B16F10 cell line than in the CT26 cell line. However, fluorescence intensity was higher in CT26 cells. Co-expression of fluorescent proteins was achieved only in a small number of cells. In conclusion, this study elucidated some of the dynamics of reporter gene expression in cancer cell lines after gene electrotransfer.

• Language: English
• Keywords: B16F10, CT26, gene electrotransfer, concomitant plasmid delivery, reporter proteins, real-time monitoring
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: MF - Faculty of Medicine; ZF - Faculty of Health Sciences
• Publication status: Published
• Publication version: Version of Record
• Year: 2023
• Number of pages: 12 str.
• Numbering: Vol. 153, art. 108490
• PID: 20.500.12556/RUL-152846
• UDC: 602.6/.7
• ISSN on article: 1878-562X
• DOI: 10.1016/j.bioelechem.2023.108490
• COBISS.SI-ID: 159032323
• Publication date in RUL: 08.12.2023

Record is a part of a journal

• Title: Bioelectrochemistry
• Publisher: Elsevier
• ISSN: 1878-562X
• COBISS.SI-ID: 23407109

Licences

• License: CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
• Link: http://creativecommons.org/licenses/by-nc-nd/4.0/
• Description: The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.

Secondary language

• Language: Slovenian
• Keywords: B16F10, CT26, elektroprenos genov

Projects

• Funder: ARRS - Slovenian Research Agency
• Project number: P3-0003
• Name: Razvoj in ovrednotenje novih terapij za zdravljenje malignih tumorjev

• Funder: ARRS - Slovenian Research Agency
• Project number: J3-2528
• Name: Imunski genski elektroprenos IL-12 in IL-2 za zdravljenje immunološko hladnih/vročih tumorjev