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Mathieu, J; Detraux, D; Kuppers, D; Wang, Y; Cavanaugh, C; Sidhu, S; Levy, S; Robitaille, A M; Ferreccio, A; Bottorff, T; McAlister, A; Somasundaram, L; Artoni, F; Battle, S; Hawkins, R D; Moon, R T; Ware, C B; Paddison, P J; Ruohola-Baker, H
Nature communications, 02/2019, Letnik: 10, Številka: 1Journal Article
To reveal how cells exit human pluripotency, we designed a CRISPR-Cas9 screen exploiting the metabolic and epigenetic differences between naïve and primed pluripotent cells. We identify the tumor suppressor, Folliculin(FLCN) as a critical gene required for the exit from human pluripotency. Here we show that FLCN Knock-out (KO) hESCs maintain the naïve pluripotent state but cannot exit the state since the critical transcription factor TFE3 remains active in the nucleus. TFE3 targets up-regulated in FLCN KO exit assay are members of Wnt pathway and ESRRB. Treatment of FLCN KO hESC with a Wnt inhibitor, but not ESRRB/FLCN double mutant, rescues the cells, allowing the exit from the naïve state. Using co-immunoprecipitation and mass spectrometry analysis we identify unique FLCN binding partners. The interactions of FLCN with components of the mTOR pathway (mTORC1 and mTORC2) reveal a mechanism of FLCN function during exit from naïve pluripotency.
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