Biopolym. Cell. 2017; 33(5):356-366.
http://dx.doi.org/10.7124/bc.00095F
Molecular and Cell Biotechnologies
Generation of HEK-293 stable cell lines with disrupted expression of ribosomal protein S6 kinase (S6K1) isoforms using the CRISPR/Cas9 genome editing system
1Zaiets I.V., 1Sivchenko A.S., 1Khoruzhenko A. I., 1Filonenko V. V.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680

Abstract

Aim. To generate HEK-293 cells with disrupted expression of S6K1 isoforms: p85, p70 and p60. Methods. CRISPR/Cas9 gene editing, Western blotting, immunofluorescent analysis, RT-PCR analysis, MTT assay, scratch assay. Results. Several clones of HEK-293 cells with a complete loss of p85/p70/p60-S6K1 protein expression were generated. The effects of p85/p70/p60-S6K1 knockout on Akt/mTORC1/S6K1 signaling and cell proliferation and migration were assessed. Conclusions. The generated cell lines can be used to study a role played by S6K1 in cell physiology and to gain more detailed insight into cellular functions of the S6K1 isoforms. The HEK-293 cells exhibit down-regulation of Akt phosphorylation on Ser473 and subsequent attenuation of cell growth rate, as well as inhibition of cell motility.
Full text: (PDF, in English)

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