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- W1531780147 abstract "This chapter outlines various techniques for analyzing X-inactivation kinetics in differentiating Embryonic stem (ES) cells. The chapter focuses particularly on changing patterns of histone modifications during X inactivation, using immunuofluorescence combined with RNA fluorescence in situ hybridization (FISH) on interphase nuclei, as well as metaphase chromosome staining combined with DNA FISH. X-chromosome inactivation provides a powerful model system to investigate the different steps in facultative heterochromatin formation. During early development, one of the two X chromosomes is transcriptionally silenced in every cell of a female embryo, thereby achieving dosage compensation between males and females for X-linked gene products. The X inactivation process is dependent on the action of a unique RNA, Xist, which coats the X chromosome in cis and induces its inactivation. Once established, the inactive state of the X chromosome is highly stable in somatic cells and is normally only reversed in the female germ line. In addition, the chapter also describes a protocol for assaying late replication timing of the inactive X chromosome. Using these techniques it is possible to determine the relative order of the following events: Xist RNA coating occurs within the first 24–48 hours of differentiation; histone H3 modifications, such as hypomethylation of Lys-4, hypoacetylation of Lys- 9, and hypermethylation of Lys-9 and Lys-27 are detectable on the X chromosome in a proportion of interphase cells as soon as Xist RNA accumulates." @default.
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- W1531780147 date "2003-01-01" @default.
- W1531780147 modified "2023-10-13" @default.
- W1531780147 title "X-Chromosome Inactivation in Mouse Embryonic Stem Cells: Analysis of Histone Modifications and Transcriptional Activity Using Immunofluorescence and FISH" @default.
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- W1531780147 doi "https://doi.org/10.1016/s0076-6879(03)76027-3" @default.
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