Tracing the transitions from pluripotency to germ cell fate with CRISPR screening

  • Nat Commun. 2018 Oct 16;9(1):4292. doi: 10.1038/s41467-018-06230-0.
Jamie A Hackett  1  2  3 ,  Yun Huang  4  5 ,  Ufuk Günesdogan  4  5  6 ,  Kristjan A Gretarsson  7 ,  Toshihiro Kobayashi  4  5  8 ,  M Azim Surani  9  10
Affiliations
  • 1. Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QN, UK. [email protected].
  • 2. Epigenetics and Neurobiology Unit, European Molecular Biology Laboratory (EMBL), via Ramarini 32, 00015, Rome, Italy. [email protected].
  • 3. Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge, CB2 3DY, UK. [email protected].
  • 4. Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QN, UK.
  • 5. Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge, CB2 3DY, UK.
  • 6. Department of Developmental Biology, University of Göttingen, Göttingen Center for Molecular Biosciences, Justus-von-Liebig Weg 11, 37077, Göttingen, Germany.
  • 7. Epigenetics and Neurobiology Unit, European Molecular Biology Laboratory (EMBL), via Ramarini 32, 00015, Rome, Italy.
  • 8. Center for Genetic Analysis of Behaviour, National Institute for Physiological Sciences, 5-1 Higashiyama Myodaiji, Okazaki, Aichi, 444-8787, Japan.
  • 9. Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QN, UK. [email protected].
  • 10. Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge, CB2 3DY, UK. [email protected].
PMID: 30327475 DOI: 10.1038/s41467-018-06230-0
Abstract

Early mammalian development entails transit through naive 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.