Rescue of Fragile X Syndrome Neurons by DNA Methylation Editing of the FMR1 Gene
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...
| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier BV
2020
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| Online Access: | https://hdl.handle.net/1721.1/126002 |
| _version_ | 1811096668769091584 |
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| author | Liu, X. Shawn Wu, Hao Krzisch, Marine Wu, Xuebing Graef, John Muffat, Julien Hnisz, Denes Li, Charles Han Yuan, Bingbing Xu, Chuanyun Li, Yun Vershkov, Dan Cacace, Angela Young, Richard A. Jaenisch, Rudolf |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Liu, X. Shawn Wu, Hao Krzisch, Marine Wu, Xuebing Graef, John Muffat, Julien Hnisz, Denes Li, Charles Han Yuan, Bingbing Xu, Chuanyun Li, Yun Vershkov, Dan Cacace, Angela Young, Richard A. Jaenisch, Rudolf |
| author_sort | Liu, X. Shawn |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T16:47:09Z |
| format | Article |
| id | mit-1721.1/126002 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T16:47:09Z |
| publishDate | 2020 |
| publisher | Elsevier BV |
| record_format | dspace |
| spelling | mit-1721.1/1260022022-10-03T08:17:57Z Rescue of Fragile X Syndrome Neurons by DNA Methylation Editing of the FMR1 Gene Liu, X. Shawn Wu, Hao Krzisch, Marine Wu, Xuebing Graef, John Muffat, Julien Hnisz, Denes Li, Charles Han Yuan, Bingbing Xu, Chuanyun Li, Yun Vershkov, Dan Cacace, Angela Young, Richard A. Jaenisch, Rudolf Massachusetts Institute of Technology. Department of Biology 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. National Institutes of Health (Grant HD045022) National Institutes of Health (Grant MH104610) National Institutes of Health (Grant R01NS088538) National Institutes of Health (Grant R01GM123511) 2020-06-26T19:45:52Z 2020-06-26T19:45:52Z 2018-02 2017-12 2019-12-10T19:23:19Z Article http://purl.org/eprint/type/JournalArticle 0092-8674 https://hdl.handle.net/1721.1/126002 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 en http://dx.doi.org/10.1016/j.cell.2018.01.012 Cell Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier BV PMC |
| spellingShingle | Liu, X. Shawn Wu, Hao Krzisch, Marine Wu, Xuebing Graef, John Muffat, Julien Hnisz, Denes Li, Charles Han Yuan, Bingbing Xu, Chuanyun Li, Yun Vershkov, Dan Cacace, Angela Young, Richard A. Jaenisch, Rudolf Rescue of Fragile X Syndrome Neurons by DNA Methylation Editing of the FMR1 Gene |
| title | Rescue of Fragile X Syndrome Neurons by DNA Methylation Editing of the FMR1 Gene |
| title_full | Rescue of Fragile X Syndrome Neurons by DNA Methylation Editing of the FMR1 Gene |
| title_fullStr | Rescue of Fragile X Syndrome Neurons by DNA Methylation Editing of the FMR1 Gene |
| title_full_unstemmed | Rescue of Fragile X Syndrome Neurons by DNA Methylation Editing of the FMR1 Gene |
| title_short | Rescue of Fragile X Syndrome Neurons by DNA Methylation Editing of the FMR1 Gene |
| title_sort | rescue of fragile x syndrome neurons by dna methylation editing of the fmr1 gene |
| url | https://hdl.handle.net/1721.1/126002 |
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