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Optimization of A549 cell transfection efficiency with a plasmid encoding the N-protein of the SARS-COV-2 virus
- Authors: Khramtsov Y.V.1, Lupanova T.N.1, Rosenkranz A.А.1,2, Georgiev G.P.1, Sobolev A.S.1,2
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Affiliations:
- Institute of Gene Biology, RAS
- Lomonosov Moscow State University Moscow
- Issue: Vol 521, No 1 (2025)
- Pages: 204-207
- Section: Articles
- URL: https://cijournal.ru/2686-7389/article/view/684007
- DOI: https://doi.org/10.31857/S2686738925020064
- ID: 684007
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Abstract
To test new antiviral drugs aimed at degrading the nucleocapsid protein (N-protein) of the SARS-CoV-2 virus, it is desirable to have cells expressing the N-protein, for which it is necessary to find conditions for the maximum achievable efficiency of cell transfection with a plasmid encoding this protein. For transfection, polyplexes were used consisting of a plasmid encoding the N-protein fused with the mRuby3 fluorescent protein and polyethyleneimine (PEI)-polyethylene glycol (PEG)-TAT peptide block copolymers. The dependence of the transfection efficiency of human lung adenocarcinoma A549 cells on the PEG/PEI and N/P ratios (the ratio of nitrogen in PEI to phosphate in DNA) was studied. Significant positive correlations were shown between transfection efficiency determined by flow cytometry, the N/P ratio, and the proportion of polyplexes sized 40–54 nm. The data obtained can serve as a basis for creating an animal model of lung cells transiently expressing the N protein of the SARS-CoV-2 virus.
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About the authors
Y. V. Khramtsov
Institute of Gene Biology, RAS
Author for correspondence.
Email: alsobolev@yandex.ru
Russian Federation, Moscow
T. N. Lupanova
Institute of Gene Biology, RAS
Email: alsobolev@yandex.ru
Russian Federation, Moscow
A. А. Rosenkranz
Institute of Gene Biology, RAS; Lomonosov Moscow State University Moscow
Email: alsobolev@yandex.ru
Russian Federation, Moscow; Moscow
G. P. Georgiev
Institute of Gene Biology, RAS
Email: alsobolev@yandex.ru
Academician of the RAS
Russian Federation, MoscowA. S. Sobolev
Institute of Gene Biology, RAS; Lomonosov Moscow State University Moscow
Email: alsobolev@yandex.ru
Corresponding Member of the RAS
Russian Federation, Moscow; MoscowReferences
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