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Optimization of A549 Cell Transfection Efficiency with a Plasmid Encoding the N-Protein of the SARS-CoV-2 Virus

  • BIOCHEMISTRY, BIOPHYSICS, AND MOLECULAR BIOLOGY
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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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ACKNOWLEDGEMENTS

The experiments were carried out using equipment of the Core Facility of the Institute of Gene Biology of the Russian Academy of Sciences.

Funding

The study was supported by a grant from the Russian Science Foundation (project no. 24-14-00170).

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Authors and Affiliations

  1. Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia

    Yu. V. Khramtsov, T. N. Lupanova, A. A. Rosenkranz,  G. P. Georgiev &  A. S. Sobolev

  2. Moscow State University, Moscow, Russia

    A. A. Rosenkranz &  A. S. Sobolev

Authors
  1. Yu. V. Khramtsov
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  2. T. N. Lupanova
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  3. A. A. Rosenkranz
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  4. G. P. Georgiev
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  5. A. S. Sobolev
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Corresponding author

Correspondence to A. S. Sobolev.

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This work does not contain any studies involving human and animal subjects.

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Translated by M. Batrukova

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Khramtsov, Y.V., Lupanova, T.N., Rosenkranz, A.A. et al. Optimization of A549 Cell Transfection Efficiency with a Plasmid Encoding the N-Protein of the SARS-CoV-2 Virus. Dokl Biochem Biophys (2025). https://doi.org/10.1134/S1607672924601136

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  • DOI: https://doi.org/10.1134/S1607672924601136

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