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

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.

Keywords

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

Journal Title

Nat Commun

Journal ISSN

2041-1723
2041-1723

Volume Title

11

Publisher

Springer Nature

Publisher DOI

https://doi.org/10.1038/s41467-020-15042-0

Rights and licensing

Except where otherwised noted, this item's license is described as Attribution 4.0 International

Sponsorship

Wellcome Trust (096738/Z/11/Z)
Medical Research Council (MR/P009948/1)

AS was funded by the 4-year Wellcome Trust PhD Scholarship; MPS was funded by a Churchill scholarship; MAS holds a Wellcome Senior Investigator Award, and grant from the MRC. The Gurdon Institute is supported through core funding from the Wellcome Trust and Cancer Research UK. WHG was supported by an EMBO long term fellowship (ALTF 263-2014). WWCT holds a Croucher Fellowship for Postdoctoral Research.

Relationships

Is derived from:

https://doi.org/10.1101/563072

Collections

University of Cambridge Research Outputs (Articles and Conferences)