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The abnormal reactivation of\n                    <jats:italic>DUX4<\/jats:italic>\n                    , an embryonic gene that is epigenetically silenced in somatic tissues, is causal to FSHD. Disease-specific reactivation of\n                    <jats:italic>DUX4<\/jats:italic>\n                    has two common characteristics, the presence of a non-canonical polyadenylation sequence within exon 3 of\n                    <jats:italic>DUX4<\/jats:italic>\n                    that stabilizes pathogenic transcripts, and the loss of repressive chromatin modifications at D4Z4, the macrosatellite repeat which encodes\n                    <jats:italic>DUX4<\/jats:italic>\n                    . We used CRISPR\/Cas9 to silence\n                    <jats:italic>DUX4<\/jats:italic>\n                    using two independent approaches. We deleted the\n                    <jats:italic>DUX4<\/jats:italic>\n                    pathogenic polyadenylation signal, which resulted in downregulation of pathogenic\n                    <jats:italic>DUX4-fl<\/jats:italic>\n                    transcripts. In another approach, we transcriptionally repressed\n                    <jats:italic>DUX4<\/jats:italic>\n                    by seeding heterochromatin using the dCas9-KRAB platform within exon 3. These feasibility of targeting\n                    <jats:italic>DUX4<\/jats:italic>\n                    experiments were initially tested in a non-myogenic carcinoma cell line that we have previously characterized. Subsequently, in an immortalized patient myoblast cell line, we demonstrated that targeting\n                    <jats:italic>DUX4<\/jats:italic>\n                    by either approach led to substantial downregulation of not only pathogenic\n                    <jats:italic>DUX4<\/jats:italic>\n                    transcripts, but also a subset of its target genes that are known biomarkers of FSHD. These findings offer proof-of-concept of the effect of silencing the polyadenylation sequence on pathogenic\n                    <jats:italic>DUX4<\/jats:italic>\n                    expression.\n                  <\/jats:p>","DOI":"10.1038\/s41598-021-92096-0","type":"journal-article","created":{"date-parts":[[2021,6,15]],"date-time":"2021-06-15T06:04:25Z","timestamp":1623737065000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":34,"title":["CRISPR mediated targeting of DUX4 distal regulatory element represses DUX4 target genes dysregulated in Facioscapulohumeral muscular dystrophy"],"prefix":"10.1038","volume":"11","author":[{"given":"Sunny","family":"Das","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian P.","family":"Chadwick","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,6,15]]},"reference":[{"key":"92096_CR1","doi-asserted-by":"publisher","first-page":"525","DOI":"10.1016\/S0960-8966(01)00201-2","volume":"11","author":"KM Flanigan","year":"2001","unstructured":"Flanigan, K. 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