Tracing the Transitions from Pluripotency to Germ Cell Fate with CRISPR Screening
Authors
Surani, A
Hackett, Jamie A
Huang, Yun
Gunesdogan, Ufuk
Holm-Gretarsson, Kristjan
Kobayashi, Toshihiro
Publication Date
2018-10-16Journal Title
Nature Communications
ISSN
2041-1723
Publisher
Springer Nature
Type
Article
Metadata
Show full item recordCitation
Surani, A., Hackett, J. A., Huang, Y., Gunesdogan, U., Holm-Gretarsson, K., & Kobayashi, T. (2018). Tracing the Transitions from Pluripotency to Germ Cell Fate with CRISPR Screening. Nature Communications https://doi.org/10.1038/s41467-018-06230-0
Abstract
Early mammalian development entails transit through naïve 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.
Keywords
Animals, Basic Helix-Loop-Helix Transcription Factors, Chromosomal Proteins, Non-Histone, Clustered Regularly Interspaced Short Palindromic Repeats, DNA-Binding Proteins, Embryonic Development, Gene Expression Regulation, Developmental, Genes, Reporter, Green Fluorescent Proteins, Mice, Mice, Transgenic, Pluripotent Stem Cells, Positive Regulatory Domain I-Binding Factor 1, Receptors, Cytoplasmic and Nuclear, Repressor Proteins, Wnt Proteins
Sponsorship
Wellcome Trust (092096/Z/10/Z)
Cancer Research Uk (None)
Embargo Lift Date
2100-01-01
Identifiers
External DOI: https://doi.org/10.1038/s41467-018-06230-0
This record's URL: https://www.repository.cam.ac.uk/handle/1810/279918
Rights
Attribution 4.0 International (CC BY)
Licence URL: http://creativecommons.org/licenses/by/4.0/
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