Screening for Factors Involved in X Chromosome Inactivation Using Haploid ESCs
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 wordsHaploid Embryonic stem cells X inactivation Genetic screening Chromatin Gene regulation
Embryonic stem cell
Phosphate buffered saline
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.