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Title: Live-cell imaging and functional dissection of Xist RNA reveal mechanisms of X chromosome inactivation and reactivation
Authors: Ha, Norbert How Ong
Lai, Lan-Tian
Chelliah, Rosi
Zhen, Yashu
Seet, Vanessa Pei Yi
Lai, Soak-Kuan
Li, Hoi-Yeung
Ludwig, Alexander
Sandin, Sara
Chen, Lingyi
Zhang, Li-Feng
Keywords: Science::Biological sciences::Molecular biology
Cell Biology
Issue Date: 2018
Source: Ha, N. H. O., Lai, L.-T., Chelliah, R., Zhen, Y., Seet, V. P. Y., Lai, S.-K., . . . Zhang, L.-F. (2018). Live-cell imaging and functional dissection of Xist RNA reveal mechanisms of X chromosome inactivation and reactivation. iScience, 8, 1-14. doi:10.1016/j.isci.2018.09.007
Series/Report no.: iScience
Abstract: We double-tagged Xist (inactivated X chromosome-specific transcript), a prototype long non-coding RNA pivotal for X chromosome inactivation (XCI), using the programmable RNA sequence binding domain of Pumilio protein, one tag for live-cell imaging and the other replacing A-repeat (a critical domain of Xist) to generate “ΔA mutant” and to tether effector proteins for dissecting Xist functionality. Based on the observation in live cells that the induced XCI in undifferentiated embryonic stem (ES) cells is counteracted by the intrinsic X chromosome reactivation (XCR), we identified Kat8 and Msl2, homologs of Drosophila dosage compensation proteins, as players involved in mammalian XCR. Furthermore, live-cell imaging revealed the obviously undersized ΔA Xist cloud signals, clarifying an issue regarding the previous RNA fluorescence in situ hybridization results. Tethering candidate proteins onto the ΔA mutant reveals the significant roles of Ythdc1, Ezh2, and SPOC (Spen) in Xist-mediated gene silencing and the significant role of Ezh2 in Xist RNA spreading.
DOI: 10.1016/j.isci.2018.09.007
Rights: © 2018 The Author(s). This is an open access article under the CC BY-NC-ND license (
Fulltext Permission: open
Fulltext Availability: With Fulltext
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