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J Vis Exp. 2019 May 22;(147). doi: 10.3791/59449.

A Non-random Mouse Model for Pharmacological Reactivation of Mecp2 on the Inactive X Chromosome.

Author information

1
Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine.
2
Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine; Department of Neuroscience, University of Virginia School of Medicine; sb5fk@virginia.edu.

Abstract

X chromosome inactivation (XCI) is the random silencing of one X chromosome in females to achieve gene dosage balance between the sexes. As a result, all females are heterozygous for X-linked gene expression. One of the key regulators of XCI is Xist, which is essential for the initiation and maintenance of XCI. Previous studies have identified 13 trans acting X chromosome inactivation factors (XCIFs) using a large-scale, loss-of-function genetic screen. Inhibition of XCIFs, such as ACVR1 and PDPK1, using short-hairpin RNA or small molecule inhibitors, reactivates X chromosome-linked genes in cultured cells. But the feasibility and tolerability of reactivating the inactive X chromosome in vivo remains to be determined. Towards this goal, a XistΔ:Mecp2/Xist:Mecp2-Gfp mouse model has been generated with non-random XCI due to deletion of Xist on one X chromosome. Using this model, the extent of inactive X reactivation was quantitated in the mouse brain following treatment with XCIF inhibitors. Recently published results show, for the first time, that pharmacological inhibition of XCIFs reactivates Mecp2 from the inactive X chromosome in cortical neurons of the living mouse brain.

PMID:
31180354
DOI:
10.3791/59449

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