Work

Specific Activation of the Expression of Growth Factor Genes in Expi293F Human Cells Using CRISPR/Cas9-SAM Technology Increases Their Proliferation

Year: 2024

Type: article

Abstract: Human cell lines play an important role in biotechnology and pharmacology. For them to grow, they need complex nutrient media containing signaling proteins - growth factors. We have tested a new approach that reduces the need of cultured human cell lines for exogenous growth factors. This approach is based on the generation of a modified cell with a selectively activated gene expression of one of the endogenous growth factors: IGF-1, FGF-2, or EIF3I. We modified the Expi293F cell line, a HEK293 cell line variant widely used in the production of recombinant proteins. Gene expression of the selected growth factors in these cells was activated using CRISPR/Cas9 technology with the synergistic activation mediators CRISPR/Cas9-SAM, which increased the expression of the selected genes at both mRNA and protein levels. Upon culturing under standard conditions, the modified lines exhibited increased proliferation. A synergistic effect was observed in co-culture of the three modified lines. In our opinion, these results indicate that this approach is promising for efficient modification of cell lines used in biotechnology.

Source: Acta Naturae

DOI: 10.32607/actanaturae.27415

Authors: P. A. Bobrovsky, E. N. Grafskaia, D. D. Kharlampieva, V. A. Manuvera, V. N. Lazarev

Institutions: Moscow Institute of Physics and Technology, Federal Research and Clinical Center of Physical-Chemical Medicine named after Y.M. Lopukhin, Federal Medical-Biological Agency

Language: English

Topic: CRISPR and Genetic Engineering

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: diamond

Funder: Russian Science Foundation