KDM6A facilitates Xist upregulation at the onset of X inactivation

Abstract

AbstractX chromosome inactivation (XCI) is a female-specific process in which one X chromosome is silenced to balance X-linked gene expression between the sexes. XCI is initiated in early development by upregulation of the lncRNAXiston the future inactive X (Xi). A subset of X-linked genes escape silencing and thus have higher expression in females, suggesting female-specific functions. One of these genes is the highly conserved geneKdm6a, which encodes a histone demethylase that removes methyl groups at H3K27 to facilitate gene expression. Here, we investigate the role of KDM6A in the regulation ofXist. We observed impaired upregulation ofXistduring early stages of differentiation in hybrid mouse ES cells following CRISPR/Cas9 knockout ofKdm6a. This is associated with reducedXistRNA coating of the Xi, suggesting diminished XCI potency. Indeed,Kdm6aknockout results in aberrant overexpression of genes from the Xi after differentiation. KDM6A binds to theXistpromoter and knockout cells show an increase in H3K27me3 atXist. These results indicate that KDM6A plays a role in the initiation of XCI through histone demethylase-dependent activation ofXistduring early differentiation.