Comparative efficiency of accessible transfection methods in model cell lines for biotechnological applications

About authors

1 Engelhardt Institute of Molecular Biology, Moscow, Russia

2 Pirogov Russian National Research Medical University, Moscow, Russia

Correspondence should be addressed: Anastasia V. Lipatova
Vavilova, 32/1, Moscow, 119991, Russia; moc.liamg@vnaavotapil

About paper

Funding: the project was supported by the Russian Science Foundation (grant number 20-75-10157 of August 14, 2020 "Research on the possibilities of obtaining recombinant strains of oncolytic viruses with tumor-specific replication and immunomodulatory protein expression").

Author contribution: Vorobyev PO, Kochetkov DV, Vasilenko KV and Lipatova AV participated equally in the laboratory experiments, preparation of the figures and interpretation of the results.

Compliance with ethical standards: the study complies with the requirements of the World Medical Association Declaration of Helsinki.

Received: 2022-04-22 Accepted: 2022-05-30 Published online: 2022-06-23

Transient gene expression is one of the most common methods in molecular biology, equally relevant for basic research projects and biotechnological industries. Despite the existence of commercial transfection systems, which afford high transfection efficiency and high expression levels of reporter genes, expanding such systems to industrial scales is often problematic due to high costs of the reagents. The well-described methods of cationic and calcium-phosphate transfection are accessible and ensure reproducible results at much lower costs. This study is aimed at comparative validation of calcium phosphate and cationic (polyethylenimine-based) transfection protocols along with the commercially available TurboFect reagent for mono- and cotransfections on a panel of commonly used cell lines including HEK293T, Huh7, BHK-21, CHO and MRC5. The efficiency of transfection with plasmid constructs encoding different fluorescent proteins was measured by flow cytometry. Of all the tested methods, calcium phosphate transfection afforded the highest efficiency of plasmid DNA delivery in all the cell lines except BHK21, for which the PEI method turned out to be more efficient than calcium phosphate transfection, and CHO, for which both methods showed comparable efficiency.

Keywords: lentiviruses, transfection, calcium phosphate transfection, polyethylenimine, cotransfection