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The majority of FXS cases are caused by transcriptional repression of the FMR1 gene due to epigenetic changes that are not recapitulated in current animal disease models. FXS patient induced pluripotent stem cell (iPSC)-derived gene edited reporter cell lines enable novel strategies to discover reactivators of FMR1 expression in human cells on a much larger scale than previously possible. Here, we describe the workflow using FXS iPSC-derived neural cell lines to conduct a massive, unbiased screen for small molecule activators of the FMR1 gene. The proof-of-principle methodology demonstrates the utility of human stem-cell-based methodology for the untargeted discovery of reactivators of the human FMR1 gene that can be applied to other diseases.<\/jats:p>","DOI":"10.3390\/cells11010069","type":"journal-article","created":{"date-parts":[[2021,12,28]],"date-time":"2021-12-28T01:18:15Z","timestamp":1640654295000},"page":"69","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["High Throughput Small Molecule Screen for Reactivation of FMR1 in Fragile X Syndrome Human Neural Cells"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5719-9028","authenticated-orcid":false,"given":"Jack F. V.","family":"Hunt","sequence":"first","affiliation":[{"name":"Waisman Center, University of Wisconsin-Madison, Madison, WI 53705, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meng","family":"Li","sequence":"additional","affiliation":[{"name":"Waisman Center, University of Wisconsin-Madison, Madison, WI 53705, USA"},{"name":"Department of Neuroscience, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, WI 53705, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6240-9843","authenticated-orcid":false,"given":"Ryan","family":"Risgaard","sequence":"additional","affiliation":[{"name":"Waisman Center, University of Wisconsin-Madison, Madison, WI 53705, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gene E.","family":"Ananiev","sequence":"additional","affiliation":[{"name":"Carbone Cancer Center Drug Discovery Core, University of Wisconsin-Madison, Madison, WI 53705, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Scott","family":"Wildman","sequence":"additional","affiliation":[{"name":"Carbone Cancer Center Drug Discovery Core, University of Wisconsin-Madison, Madison, WI 53705, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fan","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Pharmacy, University of Wisconsin-Madison, Madison, WI 53705, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4502-3084","authenticated-orcid":false,"given":"Tim S.","family":"Bugni","sequence":"additional","affiliation":[{"name":"School of Pharmacy, University of Wisconsin-Madison, Madison, WI 53705, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5128-4424","authenticated-orcid":false,"given":"Xinyu","family":"Zhao","sequence":"additional","affiliation":[{"name":"Waisman Center, University of Wisconsin-Madison, Madison, WI 53705, USA"},{"name":"Department of Neuroscience, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, WI 53705, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0360-7145","authenticated-orcid":false,"given":"Anita","family":"Bhattacharyya","sequence":"additional","affiliation":[{"name":"Waisman Center, University of Wisconsin-Madison, Madison, WI 53705, USA"},{"name":"Department of Cell and Regenerative Biology, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, WI 53705, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,12,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1016\/j.ajhg.2009.09.007","article-title":"Incidence of Fragile X Syndrome by Newborn Screening for Methylated FMR1 DNA","volume":"85","author":"Coffee","year":"2009","journal-title":"Am. 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