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Knockout of BAX and BAK1 Genes and Overexpression of BCL2, BECN1 Genes Increase Lifespan and the Maximum Density of a CHO-S Cell Culture

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Abstract

Two genome editions have resulted in the CHO cell line 4BGD with homozygous knockouts of the BAK1, BAX, DHFR and GLUL genes, and overexpression of the BCL2 and BECN1 genes. This line is capable of long-term growth without changing the culture medium and exhibits a fivefold increase in the integral cell density compared to the parental cell line under the same cultivation regime. Overexpression of the BCL2, BECN1 gene pair led to a significant increase in cell density on days 6‒10 of cultivation compared to the cell line with a knockout of the BAK1, BAX, DHFR and GLUL genes, while the integral viable cell density increases by 34%.

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Funding

The work was carried out within the framework of the state assignment of the Fundamentals of Biotechnology Federal Research Center.

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Authors and Affiliations

  1. Skryabin Institute of Bioengineering, Fundamentals of Biotechnology Federal Research Center, Russian Academy of Sciences, 117312, Moscow, Russia

    S. V. Kovnir, L. K. Dayanova, E. A. Gaiamova, L. N. Dybovsky, I. I. Vorobiev & N. A. Orlova

  2. Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997, Moscow, Russia

    L. K. Dayanova & I. I. Vorobiev

Authors
  1. S. V. Kovnir
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  2. L. K. Dayanova
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  3. E. A. Gaiamova
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  4. L. N. Dybovsky
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  5. I. I. Vorobiev
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  6. N. A. Orlova
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Correspondence to N. A. Orlova.

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Translated by I. Gordon

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Abbreviations: Bak-1, Bcl-2 antagonist/killer 1; Bax, Bcl-2-associated X factor; CHO, Chinese hamster ovary cell line; DHFR, dihydrofolate reductase; ER, endoplasmic reticulum; IVCD, integral viable cell density; gRNA, guide RNA; GS, glutamine synthetase; HT, hypoxanthine-thymidine; ORF, open reading frame.

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Kovnir, S.V., Dayanova, L.K., Gaiamova, E.A. et al. Knockout of BAX and BAK1 Genes and Overexpression of BCL2, BECN1 Genes Increase Lifespan and the Maximum Density of a CHO-S Cell Culture. Appl Biochem Microbiol 59, 1047–1052 (2023). https://doi.org/10.1134/S0003683823080057

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