Published November 28, 2025 | Version Published
Journal Article Open

Role of CEBPa in trophectoderm competence installment

  • 1. ROR icon Centre for Genomic Regulation
  • 2. ROR icon Centro Nacional de Análisis Genómico
  • 3. ROR icon University of Cambridge
  • 4. ROR icon Pompeu Fabra University
  • 5. ROR icon Institució Catalana de Recerca i Estudis Avançats
  • 6. Research and Development, Eugin Group, Barcelona 08006, Spain.
  • 7. ROR icon University of Barcelona
  • 8. ROR icon California Institute of Technology

Abstract

During mouse embryogenesis, totipotency is gradually lost, and, at the 16-cell stage, blastomeres begin to bifurcate into trophectoderm (future placenta) and inner cell mass (future fetus). Although this process is well studied, when and how blastomeres acquire the competence for lineage specification remains unclear. Here, we describe that CEBPa becomes up-regulated at the transition from the two- to the four-cell stage by NR5A2 and is also selectively expressed in the trophectoderm at the blastocyst stage. Its knockout decreases the proportion of trophectoderm cells and delays the morula to blastocyst transition. Conversely, CEBPa overexpression in mouse embryonic stem cells, used as a proxy, drives their differentiation into trophectoderm-like cells, enabling the identification of CEBPa-regulated trophectoderm-specific enhancers. A subset of these enhancers, associated with key trophectoderm-related transcription factor genes, is primed or activated in four- and eight-cell embryos. Together, our data suggest that CEBPa plays a role in the installment of trophectoderm competence before the first lineage bifurcation and in trophectoderm specification.

Copyright and License

© 2025 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License 4.0 (CC BY).

Acknowledgement

We dedicate this paper to John Gurdon whose unwavering enthusiasm and belief in our studies on embryonic heterogeneity and cell fate specification inspired and sustained our work. We thank M. de Andres and the Genomics, Flow Cytometry, and Imaging facilities of the CRG for technical assistance; B. Payer and B. Deplancke for discussions and W. Xie for sharing embryo sequencing data crucial for our study. We acknowledge support of the Spanish Ministry of Science and Innovation through the Centro de Excelencia Severo Ochoa (CEX2020-001049-S, MCIN/AEI/10.13039/501100011033), and the Generalitat de Catalunya through the CERCA programme.

Funding

Work in the laboratory of T.G. was supported by the CRG, the Spanish Ministry of Economy, Industry and Competitiveness (Plan Estatal PID2019-109354GB-I00/AEI/10.13039/501100011033), and Secretariat of Universities and Research, Ministry of Enterprise and Knowledge of the Government of Catalonia (SGR 726). Work in the laboratory of M.Z.-G. was supported by Wellcome Trust (207415/Z/17/Z) and NIH (RO1HD100456A and RO1HD100456) grants. Work in the laboratory of M.I. was supported by Spanish Ministry of Economy, Industry and Competitiveness (Plan Estatal BFU2017-89201-P). Work in the laboratory of M.I. was supported by the Spanish Ministry of Economy, Industry and Competitiveness (Plan Estatal BFU2017-89201-P), and cofunded by ERDF.

Conflict of Interest

H.H. is the cofounder and Chief Scientific Officer of Omniscope; a scientific advisory board member of Nanostring, Bruker, and MiRXES; and a consultant to Moderna and Singularity and received an honorarium from Genentech. The authors declare that they have no other competing interests.

Data Availability

Data and Materials Availability:

All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. Requests for plasmids and cell lines generated in this study should be directed to the lead contact (thomas.graf@crg.eu). For bulk RNA-seq data, view GEO accession GSE284144; ATAC-seq data, view GEO accession GSE284145; ChIP-seq data, view GEO accession GSE284146; and single-cell multiome data, view GEO accession GSE284147.

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Additional details

Identifiers

Funding

National Institutes of Health
RO1HD100456A
National Institutes of Health
RO1HD100456
Wellcome Trust
207415/Z/17/Z
Ministerio de Economía y Competitividad
Plan Estatal PID2019-109354GB-I00/AEI/10.13039/501100011033
Spanish Ministry of Economy, Industry and Competitiveness
Plan Estatal BFU2017-89201-P
Ministry of Enterprise and Knowledge of the Government of Catalonia
SGR 726

Dates

Accepted
2025-10-30
Accepted

Caltech Custom Metadata

Caltech groups
Division of Biology and Biological Engineering (BBE)
Publication Status
Published