A critical role of PRDM14 in human primordial germ cell fate revealed by inducible degrons

Nat Commun. 2020 Mar 9;11(1):1282. doi: 10.1038/s41467-020-15042-0.

Abstract

PRDM14 is a crucial regulator of mouse primordial germ cells (mPGCs), epigenetic reprogramming and pluripotency, but its role in the evolutionarily divergent regulatory network of human PGCs (hPGCs) remains unclear. Besides, a previous knockdown study indicated that PRDM14 might be dispensable for human germ cell fate. Here, we decided to use inducible degrons for a more rapid and comprehensive PRDM14 depletion. We show that PRDM14 loss results in significantly reduced specification efficiency and an aberrant transcriptome of hPGC-like cells (hPGCLCs) obtained in vitro from human embryonic stem cells (hESCs). Chromatin immunoprecipitation and transcriptomic analyses suggest that PRDM14 cooperates with TFAP2C and BLIMP1 to upregulate germ cell and pluripotency genes, while repressing WNT signalling and somatic markers. Notably, PRDM14 targets are not conserved between mouse and human, emphasising the divergent molecular mechanisms of PGC specification. The effectiveness of degrons for acute protein depletion is widely applicable in various developmental contexts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics
  • Cell Differentiation
  • Cell Lineage*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism
  • Gene Expression Regulation
  • Germ Cells / cytology*
  • Germ Cells / metabolism*
  • HEK293 Cells
  • Humans
  • Indoleacetic Acids / pharmacology
  • Mice
  • Promoter Regions, Genetic / genetics
  • Protein Binding
  • Proteolysis*
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Transcriptome / genetics

Substances

  • DNA-Binding Proteins
  • Indoleacetic Acids
  • PRDM14 protein, human
  • Prdm14 protein, mouse
  • RNA-Binding Proteins
  • Transcription Factors