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Manipulation of Xist Imprinting in Mouse Preimplantation Embryos

  • Atsushi Fukuda
  • Akihiro Umezawa
  • Hidenori Akutsu
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1861)

Abstract

X-chromosome inactivation (XCI) is established in preimplantation embryos in mice. Prior to the establishment of XCI, expression of the long noncoding RNA (lncRNA) Xist is essential. Xist expression in mouse preimplantation embryos is imprinted, and paternal Xist is predominantly expressed. Due to the manner of imprinting, maternal Xist is always silenced. However, the nature of imprinting to repress maternal Xist is variable. For example, parthenogenetic embryos that are composed of two maternal genomes exhibit maternal Xist derepression during the preimplantation phases. Maintenance of Xist imprinting to repress maternal Xist depends on the chromatin condensation states and the dosage of Rnf12, an essential Xist activator. Therefore, alterations of chromatin states and Rnf12 expression levels lead to maternal Xist derepression. In this chapter, we describe the method for derepressing maternal Xist by various approaches, such as mRNA injection, small molecule treatment, and nuclear transfer.

Key words

Imprinted XCI Maternal Xist Chromatin condensation 

Notes

Acknowledgments

This work was supported by JSPS postdoctoral fellowships for research abroad to A.F. JSPS KAKENHI Grant-in-Aid for Young Scientists (26861350) to A.F.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Atsushi Fukuda
    • 1
    • 2
  • Akihiro Umezawa
    • 1
  • Hidenori Akutsu
    • 1
  1. 1.Center for Regenerative MedicineNational Center for Child Health and DevelopmentTokyoJapan
  2. 2.Department of Stem Cell and Regenerative BiologyHarvard UniversityCambridgeUSA

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