(T1260) GENOME-WIDE GAIN-OF-FUNCTION SCREEN IDENTIFIES NUP133 AS A KEY REGULATOR of the PLURIPOTENT-TO-TOTIPOTENT TRANSITION IN MOUSE EMBRYONIC STEM CELLS

Abstract: In mice, totipotency is exclusively observed in zygotes and two-cell (2C) embryos, enabling their differentiation into both embryonic and extra-embryonic lineages. Elucidating the regulatory mechanisms underlying totipotency is essential for the stable in vitro culture of totipotent cells and for advancing our understanding of cellular plasticity during early embryonic development. In this study, we employed piggyBac (PB) transposon-mediated gene activation vectors in conjunction with mouse embryonic stem cells (mESCs) harboring Zscan4::GFP and 2C::tdTomato reporters to conduct a genome-wide gain-of-function forward genetic screen. This screen aimed to identify key factors driving the transition from pluripotency to totipotency. Our findings reveal that the nucleoporin NUP133 plays a pivotal role in facilitating this state transition. Further analysis demonstrated that the totipotent state is associated with a more mature nuclear pore complex (NPC) compared to the pluripotent state, a phenotype similarly induced by Nup133 overexpression in mESCs. These results highlight the critical role of NPCs in the regulation of totipotency and provide new insights into the molecular mechanisms governing cellular plasticity during early development.

Funding Source: The CAMS Innovation Fund for Medical Sciences (2021-I2M-1-024 and 2021-I2M-1-019); National Natural Science Foundation of China (32370898); Beijing Natural Science Foundation (5232024)

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