title: An RNA tool kit to study the status of mouse ES cells: sex determination and stemness.
abstract: Mouse embryonic stem cells (mESCs) are pluripotent stem cells derived from the inner cell mass of the blastocyst. They can be maintained under controlled culture conditions in a pluripotent state, or be induced to differentiate into all derivatives of the three primary germ layers: ectoderm, endoderm and mesoderm. Several studies have characterised the coding and non-coding (nc) RNA repertoires of mESCs, uncovering highly dynamic variations during the process of differentiation, but also qualitative differences pertaining to sex. For example, up-regulation of the long non-coding RNA Xist on the X chromosome induces gene silencing and X inactivation exclusively during female mESC differentiation. In contrast, specific small RNAs have been shown to be up-regulated during male mESC differentiation. Here, we illustrate how a small set of key coding and ncRNAs can be exploited as dynamic and sensitive markers of the stemness and/or the differentiation status of male or female mESC lines. We describe adapted techniques for the extended characterization and analysis of mESCs from as little material as that cultured in a single 75cm(2) flask.
authors.affiliation: Swiss Federal Institute of Technology Zurich, Department of Biology, Chair of RNA Biology, Zurich, Switzerland; Life Science Zurich Graduate School, Plant Sciences Program, University of Zurich, Zurich, Switzerland.
authors.forename: F
authors.initials: F
authors.lastname: Jay
authors.forename: C
authors.initials: C
authors.lastname: Ciaudo
mesh_terms: D000818:Animals; D053624:Blastocyst Inner Cell Mass / Q000166:cytology*; D002454:Cell Differentiation; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology*; D018507:Gene Expression Regulation, Developmental; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology*; D012313:RNA / Q000378:metabolism; D062085:RNA, Long Noncoding / Q000235:genetics; D022661:RNA, Untranslated / Q000235:genetics*; D019849:Sex Determination Processes / Q000235:genetics*; D049951:X Chromosome Inactivation / Q000235:genetics
chemical_list: D062085:RNA, Long Noncoding; D022661:RNA, Untranslated; C077885:XIST non-coding RNA; D012313:RNA
keywords: Non-coding RNAs; RNAs; Small RNAs; Stemness and differentiation; mESCs