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Screening for Factors Involved in X Chromosome Inactivation Using Haploid ESCs

  • Asun Monfort
  • Giulio Di Minin
  • Anton Wutz
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1861)

Abstract

Forward genetics can provide insight into molecular pathways as has been demonstrated by advances in cell biology from comprehensive genetic studies in simple organisms. Recently, techniques have become available that promise a similar potential for understanding developmental pathways in mammals. Here we describe a genetic forward screening approach for identifying factors involved in X chromosome inactivation that is based on haploid mouse embryonic stem cells. Using a genetically encoded selection system screening of large mutant pools can identify candidate silencing factors with high confidence.

Key words

Haploid Embryonic stem cells X inactivation Genetic screening Chromatin Gene regulation 

Abbreviations

ESC

Embryonic stem cell

PBS

Phosphate buffered saline

RT

Room temperature

TX100

Triton X-100

Notes

Acknowledgment

This work received support from the Swiss National Science Fond (grants 31003A_152814 and 316030_145026). GDM was supported by an ETH Postdoctoral Fellowship. We are grateful to T. Brummelkamp for reagents and advice for the viral gene trap mutagenesis and insertion identification.

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Copyright information

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

Authors and Affiliations

  1. 1.Department of Biology, Institute of Molecular Health SciencesSwiss Federal Institute of Technology, ETH HönggerbergZurichSwitzerland

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