Tracing the Transitions from Pluripotency to Germ Cell Fate with CRISPR Screening
2018
Early mammalian development entails a series of cell fate transitions that includes transit through naive pluripotency to post-implantation
epiblast. This subsequently gives rise to
primordial germ cells(PGC), the founding population of the
germlinelineage. To investigate the
gene regulatory networksthat control these critical cell fate decisions, we developed a compound-reporter system to track cellular identity in a model of PGC specification (PGC-like cells; PGCLC), and coupled it with unbiased genome-wide CRISPR screening. This enabled identification of key genes both for exit from pluripotency and for acquisition of PGC fate, with further characterisation revealing a central role for the transcription factors Nr5a2 and Zfp296 in
germlineontogeny. Abrogation of these genes results in significantly impaired PGCLC development due to widespread activation (Nr5a2-/-) or inhibition (Zfp296-/-) of WNT pathway components. This leads to aberrant upregulation of the somatic programme or failure to appropriately activate
germlinegenes in PGCLC, respectively, and consequently loss of germ cell identity. Overall our study places Zfp296 and Nr5a2 as key components of an expanded PGC
gene regulatory network, and outlines a transferable strategy for identifying critical regulators of
complex cellfate transitions.
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