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

P A Bobrovsky; E N Grafskaia; D D Kharlampieva; V A Manuvera; V N Lazarev

doi:10.32607/actanaturae.27415

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

Acta Naturae. 2024 Jul-Sep;16(3):25-37. doi: 10.32607/actanaturae.27415.

Authors

P A Bobrovsky^{1

2}, E N Grafskaia¹, D D Kharlampieva¹, V A Manuvera^{1

2}, V N Lazarev^{1

2}

Affiliations

¹ Lopukhin Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, Moscow,119435 Russian Federation.
² Moscow Institute of Physics and Technology, Moscow, 141701 Russian Federation.

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.

Keywords: CRISPR/Cas9-SAM; EIF3I; FGF-2; HEK293; IGF-1; proliferation.