1. A critical role of PRDM14 in human primordial germ cell fate revealed by inducible degrons
- Author
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Anastasiya Sybirna, Walfred W. C. Tang, Sabine Dietmann, M. Azim Surani, Wolfram H. Gruhn, Ran Brosh, Merrick Pierson Smela, Pierson Smela, Merrick [0000-0001-5816-7098], Brosh, Ran [0000-0001-9714-8623], Surani, M Azim [0000-0002-8640-4318], Apollo - University of Cambridge Repository, and Surani, M. Azim [0000-0002-8640-4318]
- Subjects
0301 basic medicine ,Germline development ,Cellular differentiation ,General Physics and Astronomy ,13 ,14 ,Transcriptome ,13/1 ,Mice ,0302 clinical medicine ,13/44 ,14/19 ,Promoter Regions, Genetic ,lcsh:Science ,health care economics and organizations ,Regulation of gene expression ,631/136/2434 ,Multidisciplinary ,article ,RNA-Binding Proteins ,Cell Differentiation ,3. Good health ,Cell biology ,13/31 ,DNA-Binding Proteins ,medicine.anatomical_structure ,38/35 ,38/77 ,Genetic techniques ,Reprogramming ,Germ cell ,Protein Binding ,Science ,education ,631/532/2064 ,38/90 ,13/106 ,13/109 ,Biology ,General Biochemistry, Genetics and Molecular Biology ,38 ,03 medical and health sciences ,Pluripotent stem cells ,13/21 ,13/100 ,medicine ,38/88 ,Animals ,Humans ,Cell Lineage ,Embryonic Stem Cells ,45/91 ,Indoleacetic Acids ,HEK 293 cells ,General Chemistry ,Embryonic stem cell ,030104 developmental biology ,Germ Cells ,HEK293 Cells ,Gene Expression Regulation ,631/1647/1513 ,Proteolysis ,lcsh:Q ,38/15 ,CRISPR-Cas Systems ,Chromatin immunoprecipitation ,030217 neurology & neurosurgery ,Transcription Factors - 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., PRDM14 is a critical transcription factor for mouse primordial germ cell specification, but its role in human remains unclear. Here, PRDM14 protein depletion using auxin-inducible degron uncovers a critical role for human germ cell specification, but regulation of a different set of target genes than in mouse.
- Published
- 2020