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De-erosion of X chromosome dosage compensation by the editing of XIST regulatory regions restores the differentiation potential in hPSCs.
- Source :
-
Cell reports methods [Cell Rep Methods] 2022 Nov 29; Vol. 2 (12), pp. 100352. Date of Electronic Publication: 2022 Nov 29 (Print Publication: 2022). - Publication Year :
- 2022
-
Abstract
- Human pluripotent stem cells (hPSCs) regularly and irreversibly show the erosion of X chromosome inactivation (XCI) by long non-coding RNA (lncRNA) XIST silencing, causing challenges in various applications of female hPSCs. Here, we report reliable methods to reactivate XIST with monoallelic expression in female hPSCs. Surprisingly, we find that the editing of XIST regulatory regions by Cas9-mediated non-homologous end joining is sufficient for the reactivation of XIST by endogenous systems. Proliferated hPSCs with XIST reactivation show XCI from an eroded X chromosome, suggesting that hPSCs with normal dosage compensation might lead to a growth advantage. Furthermore, the use of targeting vectors, including the XIST regulatory region sequences and selection cassette, enables XIST reactivation in hPSCs with high efficiency. XIST -reactivated hPSCs can show the restoration of differentiation potential. Thus, our findings demonstrate that XIST re-expression is a beneficial method to maximize the use of female hPSCs in various applications, such as proper disease modeling.<br />Competing Interests: A.F. is applying for a patent for the method to generate XIST re-expressing female hPSCs by gRNA-mediated XIST demethylation (2022-062974).<br /> (© 2022 The Author(s).)
Details
- Language :
- English
- ISSN :
- 2667-2375
- Volume :
- 2
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Cell reports methods
- Publication Type :
- Academic Journal
- Accession number :
- 36590687
- Full Text :
- https://doi.org/10.1016/j.crmeth.2022.100352