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.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Chromosomal Proteins, Non-Histone
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • DNA-Binding Proteins / genetics
  • Embryonic Development / genetics
  • Gene Expression Regulation, Developmental*
  • Genes, Reporter
  • Green Fluorescent Proteins / genetics
  • Mice
  • Mice, Transgenic
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / physiology*
  • Positive Regulatory Domain I-Binding Factor 1 / genetics
  • Receptors, Cytoplasmic and Nuclear / genetics
  • Repressor Proteins / genetics
  • Wnt Proteins / genetics


  • Basic Helix-Loop-Helix Transcription Factors
  • Chromosomal Proteins, Non-Histone
  • DNA-Binding Proteins
  • Dppa3 protein, mouse
  • Enhancer of Split Groucho protein, mouse
  • Nr5a2 protein, mouse
  • Prdm1 protein, mouse
  • Receptors, Cytoplasmic and Nuclear
  • Repressor Proteins
  • Wnt Proteins
  • zinc finger protein 296, mouse
  • Green Fluorescent Proteins
  • Positive Regulatory Domain I-Binding Factor 1