Study of the effect of PRKDC knockout on the function of ectodermal genes in mouse embryonic stem cells

Backgrounds: Stem cells are characterized by their ability to self-renew and differentiate tonumerous cell lines. PRKDC encodes the catalytic subunit of the DNA‐dependent protein kinase(DNA-PKcs) playing a key role in non-homologous end-joining pathway. Also, it has beenshown that PRKDC is modulated in disorders with mental retardation. Due to the differentiationof ectoderm to central and peripheral nervous system, we aimed to study the potential role ofPRKDC on the expression of ectoderm genes.Materials and Methods: TT۱ stem cells in the form of PRKDC-/- and wild-type were cultured indefined medium. RNA and protein were extracted and then, the expression of NOG, BMP۴ andFGF۸ was quantitatively evaluated by real-time PCR and western blotting.Results: Our data showed that FGF۸ and NOG were upregulated in PRKDC-/- cells rather thanwild-types. However, BMP۴ was downregulated in the absence of PRKDC.Conclusion: Collectively, the results demonstrate the role of PRKDC in the function ofectoderm genes.

کلیدواژه ها:

PRKDC ، CRISPR/Cas۹ ، Ectoderm ، CNS