Tracing the Transitions from Pluripotency to Germ Cell Fate with CRISPR Screening

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Hackett, Jamie A 
Huang, Yun 
Gunesdogan, Ufuk 
Holm-Gretarsson, Kristjan 

Early mammalian development entails transit through naïve pluripotency towards post-implantation epiblast, which subsequently gives rise to primordial germ cells (PGC), the founding germline population. To investigate these cell fate transitions, we developed a compound-reporter to track cellular identity in a model of PGC specification (PGC-like cells;PGCLC), and coupled it with genome-wide CRISPR-screening. We identify key genes both for exit from pluripotency and for acquisition of PGC fate, and characterise a central role for the transcription-regulators Nr5a2 and Zfp296 in germline ontogeny. Abrogation of these genes results in widespread activation (Nr5a2-/-) or inhibition (Zfp296-/-) of WNT-pathway factors in PGCLC. This leads to aberrant upregulation of the somatic programme or failure to activate germline-genes, respectively, and consequently loss of germ cell identity. 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 cell fate decisions.

Animals, Basic Helix-Loop-Helix Transcription Factors, Chromosomal Proteins, Non-Histone, Clustered Regularly Interspaced Short Palindromic Repeats, DNA-Binding Proteins, Embryonic Development, Gene Expression Regulation, Developmental, Genes, Reporter, Green Fluorescent Proteins, Mice, Mice, Transgenic, Pluripotent Stem Cells, Positive Regulatory Domain I-Binding Factor 1, Receptors, Cytoplasmic and Nuclear, Repressor Proteins, Wnt Proteins
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Nature Communications
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Springer Nature
Wellcome Trust (092096/Z/10/Z)
Cancer Research Uk (None)