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Chromosomal silencing and localization are mediated by different domains of Xist RNA

Nature Genetics volume 30, pages 167–174 (2002)Cite this article

Abstract

The gene Xist initiates the chromosomal silencing process of X inactivation in mammals. Its product, a noncoding RNA, is expressed from and specifically associates with the inactive X chromosome in female cells. Here we use an inducible Xist expression system in mouse embryonic stem cells that recapitulates long-range chromosomal silencing to elucidate which Xist RNA sequences are necessary for chromosomal association and silencing. We show that chromosomal association and spreading of Xist RNA can be functionally separated from silencing by specific mutations. Silencing requires a conserved repeat sequence located at the 5′ end of Xist. Deletion of this element results in Xist RNA that still associates with chromatin and spreads over the chromosome but does not effect transcriptional repression. Association of Xist RNA with chromatin is mediated by functionally redundant sequences that act cooperatively and are dispersed throughout the remainder of Xist but show little or no homology.

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Figure 1: Homing of single-copy Xist cDNA transgenes to the Hprt locus.
Figure 2: Summary of Xist mutations.
Figure 3: Analysis of expression and localization of mutated Xist RNAs.
Figure 4: Targeting the endogenous Xist locus in male mouse J1 ES cells.
Figure 5: Transgenic Xist expression leads to MCB formation.
Figure 6: Analysis of the 5′ repeat element of Xist.

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Acknowledgements

We are grateful for the ability to use the microscopes of the W.M. Keck Biological Imaging Facility. This work was supported by grants from the National Institutes of Health and the Max Kade Foundation, and by the Human Frontiers Science Program Organization.

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  1. Whitehead Institute for Biomedical Research, Nine Cambridge Center, Cambridge, 02142, Massachusetts, USA

    Anton Wutz, Theodore P. Rasmussen & Rudolf Jaenisch

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Correspondence to Rudolf Jaenisch.

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Wutz, A., Rasmussen, T. & Jaenisch, R. Chromosomal silencing and localization are mediated by different domains of Xist RNA. Nat Genet 30, 167–174 (2002). https://doi.org/10.1038/ng820

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