Rescue of Fragile X Syndrome Neurons by DNA Methylation Editing of the FMR1 Gene
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nihms-941632.pdf
Description
Accepted version
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3.14 MB
Format
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Checksum (MD5)
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Author(s)
Liu, X. Shawn
Wu, Hao
Krzisch, Marine
Wu, Xuebing
Graef, John
Muffat, Julien
Hnisz, Denes
Li, Charles Han
Yuan, Bingbing
Xu, Chuanyun
Date Issued
February 2018
Journal
Cell
Publisher
Elsevier BV
Citation
Liu, X.Shawn et al. "Rescue of Fragile X Syndrome Neurons by DNA Methylation Editing of the FMR1 Gene." Cell 172, 5 (February 2018): 979-992 © 2018 Elsevier Inc
Version
Author's final manuscript
Abstract
Fragile X syndrome (FXS), the most common genetic form of intellectual disability in males, is caused by silencing of the FMR1 gene associated with hypermethylation of the CGG expansion mutation in the 5′ UTR of FMR1 in FXS patients. Here, we applied recently developed DNA methylation editing tools to reverse this hypermethylation event. Targeted demethylation of the CGG expansion by dCas9-Tet1/single guide RNA (sgRNA) switched the heterochromatin status of the upstream FMR1 promoter to an active chromatin state, restoring a persistent expression of FMR1 in FXS iPSCs. Neurons derived from methylation-edited FXS iPSCs rescued the electrophysiological abnormalities and restored a wild-type phenotype upon the mutant neurons. FMR1 expression in edited neurons was maintained in vivo after engrafting into the mouse brain. Finally, demethylation of the CGG repeats in post-mitotic FXS neurons also reactivated FMR1. Our data establish that demethylation of the CGG expansion is sufficient for FMR1 reactivation, suggesting potential therapeutic strategies for FXS. Rescue of fragile X syndrome neurons by CRISPR-mediated DNA methylation editing of the FMR1 gene.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Terms of Use
Creative Commons Attribution-NonCommercial-NoDerivs License
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DOI of Published Version
https://doi.org/10.1016/j.cell.2018.01.012
