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Increase in Sensitivity of HEK293FT Cells to Influenza Infection by CRISPR-Cas9-Mediated Knockout of IRF7 Transcription Factor

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Abstract—

Interferon-regulated factors play a central role in the activation of the innate immune response. The interferon-regulatory factor 7 (IRF7) is one of the factors that are quickly activated and are involved in a cellular response to a viral infection. In this work, monoclonal lines, based on HEK293FT cells defective in the IRF7 gene, were obtained using a CRISPR-Cas9 genome-editing method. These lines differed in the viability, proliferation rate, and susceptibility to infection with influenza A virus. Transcriptomic analysis of the most susceptible cell clone revealed differential expression of IRF7 factor as well as the other interferon-regulated genes.

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ACKNOWLEDGMENTS

The research was performed using the equipment of the Interdisciplinary centre for shared use of Kazan Federal University (Kasan, Russia) and the Center for shared Use “Genomics” (Genomics Core Facility, ICBFM SB RAS, Novosibirsk, Russia).

Funding

This work was supported by the Russian Science Foundation (project no. 18-75-10069) and partially (in development of basic methods) by State Budget Project of ICBFM SB RAS АААА-А17-117020210023-1.

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Authors and Affiliations

  1. Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, 630090, Novosibirsk, Russia

    A. B. Komissarov, M. V. Sergeeva, E. V. Mozhaeva, N. V. Eshchenko, A. D. Vasilieva, S. P. Medvedev, A. A. Malakhova, E. A. Balakhonova, E. S. Zhuravlev, D. V. Semenov, V. A. Richter & G. A. Stepanov

  2. Smorodintsev Research Institute of Influenza, Ministry of Health of the Russian Federation, 197376, St. Petersburg, Russia

    A. B. Komissarov, M. V. Sergeeva, A. D. Vasilieva & K. A. Vasilyev

  3. Federal Research Center Institute of Cytology and Genetics, Siberian Branch, Russian Academy of Sciences, 630090, Novosibirsk, Russia

    S. P. Medvedev & A. A. Malakhova

  4. Institute of Fundamental Medicine and Biology, Kazan Federal University, 420008, Kazan, Russia

    S. Yu. Malanin & T. V. Grigoryeva

Authors
  1. A. B. Komissarov
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  2. M. V. Sergeeva
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  3. E. V. Mozhaeva
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  4. N. V. Eshchenko
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  5. A. D. Vasilieva
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  6. K. A. Vasilyev
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  7. S. P. Medvedev
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  8. A. A. Malakhova
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  9. E. A. Balakhonova
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  10. S. Yu. Malanin
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  11. T. V. Grigoryeva
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  12. E. S. Zhuravlev
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  13. D. V. Semenov
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  14. V. A. Richter
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  15. G. A. Stepanov
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Corresponding author

Correspondence to G. A. Stepanov.

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The authors declare that they have no conflict of interest.

Statement on the Welfare of Animals

This article does not contain any studies involving animals performed by any of the authors.

Statement of Compliance with Standards of Research involving Humans as Subjects

This article does not contain any studies involving human participants performed by any of the authors.

Additional information

Translated by A. Barkhash

Abbreviations: IRF, interferon regulatory factors; NMD, nonsense-mediated mRNA decay; CRISPR, clustered regularly interspaced short palindromic repeats; Cas9, CRISPR associated protein 9; GFP, green fluorescent protein.

Corresponding author: phone: +7 (383) 363-51-89; e-mail: stepanovga@niboch.nsc.ru.

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Komissarov, A.B., Sergeeva, M.V., Mozhaeva, E.V. et al. Increase in Sensitivity of HEK293FT Cells to Influenza Infection by CRISPR-Cas9-Mediated Knockout of IRF7 Transcription Factor. Russ J Bioorg Chem 45, 749–757 (2019). https://doi.org/10.1134/S1068162019060232

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  • DOI: https://doi.org/10.1134/S1068162019060232

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