@article{4a4d42c5f3084638bcd5db730498e4f3,
title = "Klf5 establishes bi-potential cell fate by dual regulation of ICM and TE specification genes",
abstract = "Early blastomeres of mouse preimplantation embryos exhibit bi-potential cell fate, capable of generating both embryonic and extra-embryonic lineages in blastocysts. Here we identify three major two-cell-stage (2C)-specific endogenous retroviruses (ERVs) as the molecular hallmark of this bi-potential plasticity. Using the long terminal repeats (LTRs) of all three 2C-specific ERVs, we identify Kr{\"u}ppel-like factor 5 (Klf5) as their major upstream regulator. Klf5 is essential for bi-potential cell fate; a single Klf5-overexpressing embryonic stem cell (ESC) generates terminally differentiated embryonic and extra-embryonic lineages in chimeric embryos, and Klf5 directly induces inner cell mass (ICM) and trophectoderm (TE) specification genes. Intriguingly, Klf5 and Klf4 act redundantly during ICM specification, whereas Klf5 deficiency alone impairs TE specification. Klf5 is regulated by multiple 2C-specific transcription factors, particularly Dux, and the Dux/Klf5 axis is evolutionarily conserved. The 2C-specific transcription program converges on Klf5 to establish bi-potential cell fate, enabling a cell state with dual activation of ICM and TE genes.",
keywords = "ICM, Klf4, Klf5, MERVL, ORR1A0, ORR1A1, TE, preimplantation development",
author = "Martin Kinisu and Choi, {Yong Jin} and Claudia Cattoglio and Ke Liu and {Roux de Bezieux}, Hector and Raeline Valbuena and Nicole Pum and Sandrine Dudoit and Haiyan Huang and Zhenyu Xuan and Kim, {Sang Yong} and Lin He",
note = "Funding Information: We thank Nina Xiong, Jessica Mar, Andrew Modzelewski, Joseph Martin, Pratishtha Rawat, and Suifang Mao for technical assistance, discussions, input, and critical evaluation of the assumptions and evidence provided in this study. Further, we express our deepest gratitude to Alberto de Laco and the Trono lab for their generosity in sharing Dux knockout and control ESC lines. This study utilized Berkeley{\textquoteright}s Vincent J. Coates Genomics Sequencing Laboratory at UC Berkeley (supported by the NIH S10 OD018174 instrumentation grant) and computational resources provided by Berkeley Statistics, Biostats, and UT-Dallas. L.H. is a Thomas and Stacey Siebel Distinguished Chair Professor and Bakar fellow and is supported by a Howard Hughes Medical Institute (HHMI) faculty scholar award and several grants from the NIH ( 1R21HD088885 , GRANT12095758 , and 1R21OD027053-01 ). M.K. is supported by a CRCC pre-doctoral fellowship. Funding Information: We thank Nina Xiong, Jessica Mar, Andrew Modzelewski, Joseph Martin, Pratishtha Rawat, and Suifang Mao for technical assistance, discussions, input, and critical evaluation of the assumptions and evidence provided in this study. Further, we express our deepest gratitude to Alberto de Laco and the Trono lab for their generosity in sharing Dux knockout and control ESC lines. This study utilized Berkeley's Vincent J. Coates Genomics Sequencing Laboratory at UC Berkeley (supported by the NIH S10 OD018174 instrumentation grant) and computational resources provided by Berkeley Statistics, Biostats, and UT-Dallas. L.H. is a Thomas and Stacey Siebel Distinguished Chair Professor and Bakar fellow and is supported by a Howard Hughes Medical Institute (HHMI) faculty scholar award and several grants from the NIH (1R21HD088885, GRANT12095758, and 1R21OD027053-01). M.K. is supported by a CRCC pre-doctoral fellowship. M.K. conceived, designed, and executed the majority of the experiments reported in Figures 1, 3, and 4. Y.J.C. performed most experiments reported in Figure 2. M.K. K.L. H.R.d.B. and Z.X. provided bioinformatic analyses on preimplantation single-cell RNA-seq analysis, motif enrichment analysis and visualization, ChIP-seq analysis and visualization, and ingenuity pathway enrichment analysis. C.C. prepared ChIP-seq samples and M.K. the MERVL-reporter. ESC data were generated by N.P. Teratoma experiments and analyses were done by Y.J.C. R.V. and M.K. Morula injections to generate chimeric blastocysts as well as all imaging and quantification were performed by S.Y.K. Y.J.C. and M.K. Depletion experiments by RNAi and CRISPR-EZ as well as downstream qPCR and immunofluorescence were done by M.K. Data mining of Cistrome DB was done by K.L. and M.K. L.H. supported the conception and design of experiments as well as manuscript writing. S.D. and H.H. proofread the prepared manuscript. The authors declare no competing interests. Publisher Copyright: {\textcopyright} 2021 The Authors",
year = "2021",
month = nov,
day = "9",
doi = "10.1016/j.celrep.2021.109982",
language = "English (US)",
volume = "37",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "6",
}