Inducible XIST-dependent X-chromosome inactivation in human somatic cells is reversible
Department of Cell Biology
Chromatin Immunoprecipitation; Chromosomes, Human, X; DNA Methylation; DNA, Complementary; *Dosage Compensation, Genetic; Doxycycline; Fibrosarcoma; Gene Silencing; Heterochromatin; Histones; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Models, Genetic; *RNA, Untranslated; Sequence Analysis, DNA; *X Chromosome Inactivation
During embryogenesis, the XIST RNA is expressed from and localizes to one X chromosome in females and induces chromosome-wide silencing. Although many changes to inactive X heterochromatin are known, the functional relationships between different modifications are not well understood, and studies of the initiation of X-inactivation have been largely confined to mouse. We now present a model system for human XIST RNA function in which induction of an XIST cDNA in somatic cells results in localized XIST RNA and transcriptional silencing. Chromatin immunoprecipitation and immunohistochemistry shows that this silencing need only be accompanied by a subset of heterochromatic marks and that these can differ between integration sites. Surprisingly, silencing is XIST-dependent, remaining reversible over extended periods. Deletion analysis demonstrates that the first exon of human XIST is sufficient for both transcript localization and the induction of silencing and that, unlike the situation in mice, the conserved repeat region is essential for both functions. In addition to providing mechanistic insights into chromosome regulation and formation of facultative heterochromatin, this work provides a tractable model system for the study of chromosome silencing and suggests key differences from mouse embryonic X-inactivation.
DOI of Published Version
Proc Natl Acad Sci U S A. 2007 Jun 12;104(24):10104-9. Epub 2007 May 30. Link to article on publisher's site
Proceedings of the National Academy of Sciences of the United States of America
Chow, Jennifer C.; Hall, Lisa L.; Baldry, Sarah E.L.; Thorogood, Nancy P.; Lawrence, Jeanne B.; and Brown, Carolyn J., "Inducible XIST-dependent X-chromosome inactivation in human somatic cells is reversible" (2007). Lawrence Lab Publications. 5.