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Identification of Spen as a Crucial Factor for Xist Function through Forward Genetic Screening in Haploid Embryonic Stem Cells
A. Monfort, G. Di Minin, A. Postlmayr, R. Freimann, F. Arieti, S. Thore, A. Wutz,
Language English Country United States
Document type Journal Article, Research Support, Non-U.S. Gov't
NLK
Cell Press Free Archives
from 2012
Directory of Open Access Journals
from 2012
Free Medical Journals
from 2012
Freely Accessible Science Journals
from 2012-01-26
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from 2012-01-26
Open Access Digital Library
from 2012-01-01
- MeSH
- Embryonic Stem Cells metabolism MeSH
- Haploidy MeSH
- Nuclear Proteins genetics metabolism MeSH
- Cells, Cultured MeSH
- Mice MeSH
- Polycomb Repressive Complex 2 genetics metabolism MeSH
- RNA, Long Noncoding genetics MeSH
- Gene Silencing * MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
In mammals, the noncoding Xist RNA triggers transcriptional silencing of one of the two X chromosomes in female cells. Here, we report a genetic screen for silencing factors in X chromosome inactivation using haploid mouse embryonic stem cells (ESCs) that carry an engineered selectable reporter system. This system was able to identify several candidate factors that are genetically required for chromosomal repression by Xist. Among the list of candidates, we identify the RNA-binding protein Spen, the homolog of split ends. Independent validation through gene deletion in ESCs confirms that Spen is required for gene repression by Xist. However, Spen is not required for Xist RNA localization and the recruitment of chromatin modifications, including Polycomb protein Ezh2. The identification of Spen opens avenues for further investigation into the gene-silencing pathway of Xist and shows the usefulness of haploid ESCs for genetic screening of epigenetic pathways.
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