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Genetic Exploration of the Exit from Self-Renewal Using Haploid Embryonic Stem Cells

Self-renewal circuitry in embryonic stem cells (ESCs) is increasingly defined. How the robust pluripotency program is dissolved to enable fate transition is less appreciated. Here we develop a forward genetic approach using haploid ESCs. We created libraries of transposon integrations and screened f...

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Detalles Bibliográficos
Autores principales: Leeb, Martin, Dietmann, Sabine, Paramor, Maike, Niwa, Hitoshi, Smith, Austin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cell Press 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3995090/
https://www.ncbi.nlm.nih.gov/pubmed/24412312
http://dx.doi.org/10.1016/j.stem.2013.12.008
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author Leeb, Martin
Dietmann, Sabine
Paramor, Maike
Niwa, Hitoshi
Smith, Austin
author_facet Leeb, Martin
Dietmann, Sabine
Paramor, Maike
Niwa, Hitoshi
Smith, Austin
author_sort Leeb, Martin
collection PubMed
description Self-renewal circuitry in embryonic stem cells (ESCs) is increasingly defined. How the robust pluripotency program is dissolved to enable fate transition is less appreciated. Here we develop a forward genetic approach using haploid ESCs. We created libraries of transposon integrations and screened for persistent self-renewal in differentiation-permissive culture. This yielded multiple mutants in the Fgf/Erk and GSK3/Tcf3 modules known to drive differentiation and in epigenetic modifiers implicated in lineage commitment. We also identified and validated factors not previously considered. These include the conserved small zinc finger protein Zfp706 and the RNA binding protein Pum1. Pum1 targets several mRNAs for naive pluripotency transcription factors and accelerates their downregulation at the onset of differentiation. These findings indicate that the dismantling of pluripotent circuitry proceeds at multiple levels. More broadly they exemplify the power of haploid ESCs for genetic interrogation of developmental processes.
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spelling pubmed-39950902014-04-24 Genetic Exploration of the Exit from Self-Renewal Using Haploid Embryonic Stem Cells Leeb, Martin Dietmann, Sabine Paramor, Maike Niwa, Hitoshi Smith, Austin Cell Stem Cell Short Article Self-renewal circuitry in embryonic stem cells (ESCs) is increasingly defined. How the robust pluripotency program is dissolved to enable fate transition is less appreciated. Here we develop a forward genetic approach using haploid ESCs. We created libraries of transposon integrations and screened for persistent self-renewal in differentiation-permissive culture. This yielded multiple mutants in the Fgf/Erk and GSK3/Tcf3 modules known to drive differentiation and in epigenetic modifiers implicated in lineage commitment. We also identified and validated factors not previously considered. These include the conserved small zinc finger protein Zfp706 and the RNA binding protein Pum1. Pum1 targets several mRNAs for naive pluripotency transcription factors and accelerates their downregulation at the onset of differentiation. These findings indicate that the dismantling of pluripotent circuitry proceeds at multiple levels. More broadly they exemplify the power of haploid ESCs for genetic interrogation of developmental processes. Cell Press 2014-03-06 /pmc/articles/PMC3995090/ /pubmed/24412312 http://dx.doi.org/10.1016/j.stem.2013.12.008 Text en © 2014 The Authors http://creativecommons.org/licenses/by/3.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Short Article
Leeb, Martin
Dietmann, Sabine
Paramor, Maike
Niwa, Hitoshi
Smith, Austin
Genetic Exploration of the Exit from Self-Renewal Using Haploid Embryonic Stem Cells
title Genetic Exploration of the Exit from Self-Renewal Using Haploid Embryonic Stem Cells
title_full Genetic Exploration of the Exit from Self-Renewal Using Haploid Embryonic Stem Cells
title_fullStr Genetic Exploration of the Exit from Self-Renewal Using Haploid Embryonic Stem Cells
title_full_unstemmed Genetic Exploration of the Exit from Self-Renewal Using Haploid Embryonic Stem Cells
title_short Genetic Exploration of the Exit from Self-Renewal Using Haploid Embryonic Stem Cells
title_sort genetic exploration of the exit from self-renewal using haploid embryonic stem cells
topic Short Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3995090/
https://www.ncbi.nlm.nih.gov/pubmed/24412312
http://dx.doi.org/10.1016/j.stem.2013.12.008
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