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  • Multi-omics approach reveals posttranscriptionally regulated genes are essential for human pluripotent stem cells.

Multi-omics approach reveals posttranscriptionally regulated genes are essential for human pluripotent stem cells.

iScience (2022-05-17)
Mio Iwasaki, Yuka Kawahara, Chikako Okubo, Tatsuya Yamakawa, Michiko Nakamura, Tsuyoshi Tabata, Yohei Nishi, Megumi Narita, Akira Ohta, Hirohide Saito, Takuya Yamamoto, Masato Nakagawa, Shinya Yamanaka, Kazutoshi Takahashi
ABSTRACT

The effects of transcription factors on the maintenance and differentiation of human-induced or embryonic pluripotent stem cells (iPSCs/ESCs) have been well studied. However, the importance of posttranscriptional regulatory mechanisms, which cause the quantitative dissociation of mRNA and protein expression, has not been explored in detail. Here, by combining transcriptome and proteome profiling, we identified 228 posttranscriptionally regulated genes with strict upregulation of the protein level in iPSCs/ESCs. Among them, we found 84 genes were vital for the survival of iPSCs and HDFs, including 20 genes that were specifically necessary for iPSC survival. These 20 proteins were upregulated only in iPSCs/ESCs and not in differentiated cells derived from the three germ layers. Although there are still unknown mechanisms that downregulate protein levels in HDFs, these results reveal that posttranscriptionally regulated genes have a crucial role in iPSC survival.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Y-27632 dihydrochloride, ≥98% (HPLC)
Roche
cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail, Protease Inhibitor Cocktail Tablets provided in a glass vial, Tablets provided in a glass vial
Sigma-Aldrich
MG-132, InSolution, ≥98%, 10 mM, reversible proteasome inhibitor
Sigma-Aldrich
Wortmannin, InSolution, ≥95%, fungal metabolite that acts as a selective, cell-permeable and irreversible inhibitor of phosphatidylinositol 3-kinase (PI 3-kinase)