{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T09:00:28Z","timestamp":1791536428365,"version":"4.3.4"},"reference-count":137,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,3,12]],"date-time":"2021-03-12T00:00:00Z","timestamp":1615507200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000223","name":"Muscular Dystrophy Canada","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100000223","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000024","name":"Canadian Institutes of Health Research","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100009192","name":"Alberta Innovates","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100009192","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010090","name":"Women and Children\u2019s Health Research Institute","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100010090","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Pharmacol."],"abstract":"<jats:p>Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant disorder characterized by progressive, asymmetric muscle weakness at the face, shoulders, and upper limbs, which spreads to the lower body with age. It is the third most common inherited muscular disorder worldwide. Around 20% of patients are wheelchair-bound, and some present with extramuscular manifestations. FSHD is caused by aberrant expression of the <jats:italic>double homeobox protein 4<\/jats:italic> (<jats:italic>DUX4<\/jats:italic>) gene in muscle. <jats:italic>DUX4<\/jats:italic> codes for a transcription factor which, in skeletal muscle, dysregulates numerous signaling activities that culminate in cytotoxicity. Potential treatments for FSHD therefore aim to reduce the expression of <jats:italic>DUX4<\/jats:italic> or the activity of its toxic protein product. In this article, we review how genetic approaches such as those based on oligonucleotide and genome editing technologies have been developed to achieve these goals. We also outline the challenges these therapies are facing on the road to translation, and discuss possible solutions and future directions<\/jats:p>","DOI":"10.3389\/fphar.2021.642858","type":"journal-article","created":{"date-parts":[[2021,3,12]],"date-time":"2021-03-12T16:09:43Z","timestamp":1615565383000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":11,"title":["Genetic Approaches for the Treatment of Facioscapulohumeral Muscular Dystrophy"],"prefix":"10.3389","volume":"12","author":[{"given":"Kenji Rowel Q.","family":"Lim","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toshifumi","family":"Yokota","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,3,12]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1533","DOI":"10.1038\/nbt1042","article-title":"Antisense and siRNA as agonists of Toll-like receptors","volume":"22","author":"Agrawal","year":"2004","journal-title":"Nat. Biotechnol."},{"key":"B2","doi-asserted-by":"publisher","first-page":"1211","DOI":"10.1261\/rna.070177.118","article-title":"RNAscope in situ hybridization-based method for detecting DUX4 RNA expression in vitro","volume":"25","author":"Amini Chermahini","year":"2019","journal-title":"RNA"},{"key":"B3","doi-asserted-by":"publisher","first-page":"93","DOI":"10.3390\/genes8030093","article-title":"Antisense oligonucleotides used to target the DUX4 mRNA as therapeutic approaches in FaciosScapuloHumeral muscular dystrophy (FSHD)","volume":"8","author":"Ansseau","year":"2017","journal-title":"Genes"},{"key":"B4","doi-asserted-by":"publisher","first-page":"S39","DOI":"10.1002\/mus.880181309","article-title":"The FSHD-linked locus D4F104S1 (p13E-11) on 4q35 has a homologue on 10qter","volume":"2","author":"Bakker","year":"1995","journal-title":"Muscle Nerve Suppl."},{"key":"B5","doi-asserted-by":"publisher","first-page":"168","DOI":"10.5582\/irdr.2016.01056","article-title":"Targeting mRNA for the treatment of facioscapulohumeral muscular dystrophy","volume":"5","author":"Bao","year":"2016","journal-title":"Intractable Rare Dis. Res."},{"key":"B6","doi-asserted-by":"publisher","first-page":"5575","DOI":"10.3390\/ijms21155575","article-title":"Membrane repair deficit in facioscapulohumeral muscular dystrophy","volume":"21","author":"Bittel","year":"2020","journal-title":"Int. J. Mol. Sci."},{"key":"B7","doi-asserted-by":"publisher","first-page":"2055","DOI":"10.1038\/mt.2011.153","article-title":"AAV6-mediated systemic shRNA delivery reverses disease in a mouse model of facioscapulohumeral muscular dystrophy","volume":"19","author":"Bortolanza","year":"2011","journal-title":"Mol. Ther."},{"key":"B8","doi-asserted-by":"publisher","first-page":"550","DOI":"10.1038\/s41467-017-00730-1","article-title":"Muscle pathology from stochastic low level DUX4 expression in an FSHD mouse model","volume":"8","author":"Bosnakovski","year":"","journal-title":"Nat. Commun."},{"key":"B9","doi-asserted-by":"publisher","first-page":"3685","DOI":"10.1242\/jcs.205427","article-title":"The DUX4 homeodomains mediate inhibition of myogenesis and are functionally exchangeable with the Pax7 homeodomain","volume":"130","author":"Bosnakovski","year":"","journal-title":"J. Cel Sci."},{"key":"B10","doi-asserted-by":"publisher","first-page":"2465","DOI":"10.1172\/JCI133303","article-title":"Transcriptional and cytopathological hallmarks of FSHD in chronic DUX4-expressing mice","volume":"130","author":"Bosnakovski","year":"2020","journal-title":"J. Clin. Invest."},{"key":"B11","doi-asserted-by":"publisher","first-page":"2766","DOI":"10.1038\/emboj.2008.201","article-title":"An isogenetic myoblast expression screen identifies DUX4-mediated FSHD-associated molecular pathologies","volume":"27","author":"Bosnakovski","year":"2008","journal-title":"EMBO J."},{"key":"B12","doi-asserted-by":"publisher","first-page":"635","DOI":"10.1097\/WCO.0000000000000849","article-title":"The prospects of targeting DUX4 in facioscapulohumeral muscular dystrophy","volume":"33","author":"Bouwman","year":"2020","journal-title":"Curr. Opin. Neurol."},{"key":"B13","doi-asserted-by":"publisher","first-page":"26801","DOI":"10.1016\/S0021-9258(18)47090-1","article-title":"Effect of phosphorothioate modification of oligodeoxynucleotides on specific protein binding","volume":"269","author":"Brown","year":"1994","journal-title":"J. Biol. Chem."},{"key":"B14","doi-asserted-by":"publisher","first-page":"4882","DOI":"10.1093\/nar\/gku142","article-title":"Sequence motifs associated with hepatotoxicity of locked nucleic acid--modified antisense oligonucleotides","volume":"42","author":"Burdick","year":"2014","journal-title":"Nucleic Acids Res."},{"key":"B15","doi-asserted-by":"publisher","first-page":"1405","DOI":"10.1038\/mt.2016.111","article-title":"Morpholino-mediated knockdown of DUX4 toward facioscapulohumeral muscular dystrophy therapeutics","volume":"24","author":"Chen","year":"","journal-title":"Mol. Ther."},{"key":"B16","doi-asserted-by":"publisher","first-page":"1508","DOI":"10.1038\/mt.2016.172","article-title":"A self-restricted CRISPR system to reduce off-target effects","volume":"24","author":"Chen","year":"","journal-title":"Mol. Ther."},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1002\/wsbm.1408","article-title":"Immunity to CRISPR Cas9 and Cas12a therapeutics","volume":"10","author":"Chew","year":"2018","journal-title":"Wiley Interdiscip. Rev. Syst. Biol. Med."},{"key":"B18","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1016\/j.molmed.2020.09.008","article-title":"Therapeutic approaches in facioscapulohumeral muscular dystrophy","volume":"27","author":"Cohen","year":"2020","journal-title":"Trends Mol. Med."},{"key":"B19","doi-asserted-by":"publisher","first-page":"3497","DOI":"10.1038\/s41467-018-05843-9","article-title":"Cas9 immunity creates challenges for CRISPR gene editing therapies","volume":"9","author":"Crudele","year":"2018","journal-title":"Nat. Commun."},{"key":"B20","doi-asserted-by":"publisher","first-page":"e0160022","DOI":"10.1371\/journal.pone.0160022","article-title":"Influence of repressive histone and DNA methylation upon D4Z4 transcription in non-myogenic cells","volume":"11","author":"Das","year":"2016","journal-title":"PLoS One"},{"key":"B21","doi-asserted-by":"publisher","first-page":"716","DOI":"10.1093\/hmg\/ddx437","article-title":"Smchd1 haploinsufficiency exacerbates the phenotype of a transgenic FSHD1 mouse model","volume":"27","author":"de Greef","year":"2018","journal-title":"Hum. Mol. Genet."},{"key":"B22","doi-asserted-by":"publisher","first-page":"1548","DOI":"10.1212\/WNL.0b013e3181f96175","article-title":"Clinical features of facioscapulohumeral muscular dystrophy 2","volume":"75","author":"de Greef","year":"2010","journal-title":"Neurology"},{"key":"B23","doi-asserted-by":"publisher","first-page":"941","DOI":"10.1038\/ng.3858","article-title":"DUX-family transcription factors regulate zygotic genome activation in placental mammals","volume":"49","author":"De Iaco","year":"2017","journal-title":"Nat. Genet."},{"key":"B24","doi-asserted-by":"publisher","first-page":"155","DOI":"10.1159\/000472291","article-title":"Physical mapping evidence for a duplicated region on chromosome 10qter showing high homology with the facioscapulohumeral muscular dystrophy locus on chromosome 4qter","volume":"3","author":"Deidda","year":"1995","journal-title":"Eur. J. Hum. Genet."},{"key":"B25","doi-asserted-by":"publisher","first-page":"7020295","DOI":"10.1155\/2017\/7020295","article-title":"Are antioxidants a potential therapy for FSHD? A review of the literature","volume":"2017","author":"Denny","year":"2017","journal-title":"Oxid. Med. Cel. Longev."},{"key":"B26","doi-asserted-by":"publisher","first-page":"11301","DOI":"10.1038\/s41598-020-68135-7","article-title":"Induction of a local muscular dystrophy using electroporation in vivo: an easy tool for screening therapeutics","volume":"10","author":"Derenne","year":"2020","journal-title":"Sci. Rep."},{"key":"B27","doi-asserted-by":"publisher","first-page":"dmm046904","DOI":"10.1242\/dmm.046904","article-title":"Cellular and animal models for facioscapulohumeral muscular dystrophy","volume":"13","author":"DeSimone","year":"2020","journal-title":"Dis. Model. Mech."},{"key":"B28","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1016\/j.omtn.2017.11.004","article-title":"A sensitive in vitro approach to assess the hybridization-dependent toxic potential of high affinity gapmer oligonucleotides","volume":"10","author":"Dieckmann","year":"2018","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"B29","doi-asserted-by":"publisher","first-page":"18157","DOI":"10.1073\/pnas.0708659104","article-title":"DUX4, a candidate gene of facioscapulohumeral muscular dystrophy, encodes a transcriptional activator of PITX1","volume":"104","author":"Dixit","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"B30","doi-asserted-by":"publisher","first-page":"244","DOI":"10.1016\/j.freeradbiomed.2016.08.007","article-title":"DUX4-induced constitutive DNA damage and oxidative stress contribute to aberrant differentiation of myoblasts from FSHD patients","volume":"99","author":"Dmitriev","year":"2016","journal-title":"Free Radic. Biol. Med."},{"key":"B31","doi-asserted-by":"publisher","first-page":"1183","DOI":"10.1242\/jcs.066399","article-title":"Breaking down the barriers: siRNA delivery and endosome escape","volume":"123","author":"Dominska","year":"2010","journal-title":"J. Cel Sci."},{"key":"B32","doi-asserted-by":"publisher","first-page":"2561","DOI":"10.1016\/j.ymthe.2017.07.014","article-title":"Quantitative antisense screening and optimization for exon 51 skipping in Duchenne muscular dystrophy","volume":"25","author":"Echigoya","year":"2017","journal-title":"Mol. Ther."},{"key":"B33","doi-asserted-by":"publisher","first-page":"17113","DOI":"10.1038\/s41598-017-17180-w","article-title":"CRISPR\/Cas9 delivery with one single adenoviral vector devoid of all viral genes","volume":"7","author":"Ehrke-Schulz","year":"2017","journal-title":"Sci. Rep."},{"key":"B34","doi-asserted-by":"publisher","first-page":"1842","DOI":"10.1038\/s41467-019-09693-x","article-title":"Multifunctional CRISPR-Cas9 with engineered immunosilenced human T cell epitopes","volume":"10","author":"Ferdosi","year":"2019","journal-title":"Nat. Commun."},{"key":"B35","doi-asserted-by":"publisher","first-page":"1256","DOI":"10.1093\/hmg\/ddu536","article-title":"Direct interplay between two candidate genes in FSHD muscular dystrophy","volume":"24","author":"Ferri","year":"2015","journal-title":"Hum. Mol. Genet."},{"key":"B36","doi-asserted-by":"publisher","first-page":"631","DOI":"10.1093\/brain\/110.3.631","article-title":"Retinal vascular abnormalities in facioscapulohumeral muscular dystrophy. A general association with genetic and therapeutic implications","volume":"110","author":"Fitzsimons","year":"1987","journal-title":"Brain"},{"key":"B37","doi-asserted-by":"publisher","first-page":"78","DOI":"10.1177\/0192623314551840","article-title":"Antisense oligonucleotide therapies: the promise and the challenges from a toxicologic pathologist\u2019s perspective","volume":"43","author":"Frazier","year":"2015","journal-title":"Toxicol. Pathol."},{"key":"B38","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1016\/s0378-1119(99)00267-x","article-title":"Nucleotide sequence of the partially deleted D4Z4 locus in a patient with FSHD identifies a putative gene within each 3.3 kb element","volume":"236","author":"Gabri\u00ebls","year":"1999","journal-title":"Gene"},{"key":"B39","doi-asserted-by":"publisher","first-page":"125","DOI":"10.1089\/hyb.2010.0094","article-title":"Immunodetection of human double homeobox 4","volume":"30","author":"Geng","year":"2011","journal-title":"Hybridoma"},{"key":"B40","doi-asserted-by":"publisher","first-page":"e123538","DOI":"10.1172\/jci.insight.123538","article-title":"AAV-mediated follistatin gene therapy improves functional outcomes in the TIC-DUX4 mouse model of FSHD","volume":"3","author":"Giesige","year":"2018","journal-title":"JCI Insight"},{"key":"B41","doi-asserted-by":"publisher","first-page":"828","DOI":"10.1136\/jmedgenet-2019-106402","article-title":"Intronic SMCHD1 variants in FSHD: testing the potential for CRISPR-Cas9 genome editing","volume":"56","author":"Goossens","year":"2019","journal-title":"J. Med. Genet."},{"key":"B42","doi-asserted-by":"publisher","first-page":"1077","DOI":"10.1016\/j.nmd.2017.09.007","article-title":"Early onset facioscapulohumeral dystrophy\u2014a systematic review using individual patient data","volume":"27","author":"Goselink","year":"2017","journal-title":"Neuromuscul. Disord."},{"key":"B43","doi-asserted-by":"publisher","first-page":"302","DOI":"10.1089\/nat.2013.0436","article-title":"Hepatotoxic potential of therapeutic oligonucleotides can Be predicted from their sequence and modification pattern","volume":"23","author":"Hagedorn","year":"2013","journal-title":"Nucleic Acid Ther."},{"key":"B44","doi-asserted-by":"publisher","first-page":"e2441","DOI":"10.1212\/WNL.0000000000009617","article-title":"Homozygous nonsense variant in LRIF1 associated with facioscapulohumeral muscular dystrophy","volume":"94","author":"Hamanaka","year":"2020","journal-title":"Neurology"},{"key":"B45","doi-asserted-by":"publisher","first-page":"863","DOI":"10.1007\/s13311-018-00675-3","article-title":"Facioscapulohumeral muscular dystrophy: update on pathogenesis and future treatments","volume":"15","author":"Hamel","year":"2018","journal-title":"Neurotherapeutics"},{"key":"B46","doi-asserted-by":"publisher","first-page":"1056","DOI":"10.1016\/j.csbj.2019.07.008","article-title":"In-silico design of DNA oligonucleotides: challenges and approaches","volume":"17","author":"Hendling","year":"2019","journal-title":"Comput. Struct. Biotechnol. J."},{"key":"B47","doi-asserted-by":"publisher","first-page":"925","DOI":"10.1038\/ng.3844","article-title":"Conserved roles of mouse DUX and human DUX4 in activating cleavage-stage genes and MERVL\/HERVL retrotransposons","volume":"49","author":"Hendrickson","year":"2017","journal-title":"Nat. Genet."},{"key":"B48","doi-asserted-by":"publisher","first-page":"R17","DOI":"10.1093\/hmg\/ddv237","article-title":"Loss of epigenetic silencing of the DUX4 transcription factor gene in facioscapulohumeral muscular dystrophy","volume":"24","author":"Hewitt","year":"2015","journal-title":"Hum. Mol. Genet."},{"key":"B49","doi-asserted-by":"publisher","first-page":"1287","DOI":"10.1093\/hmg\/3.8.1287","article-title":"Analysis of the tandem repeat locus D4Z4 associated with facioscapulohumeral muscular dystrophy","volume":"3","author":"Hewitt","year":"1994","journal-title":"Hum. Mol. Genet."},{"key":"B50","doi-asserted-by":"publisher","first-page":"1942","DOI":"10.1128\/MCB.00149-14","article-title":"Myogenic enhancers regulate expression of the facioscapulohumeral muscular dystrophy-associated DUX4 gene","volume":"34","author":"Himeda","year":"2014","journal-title":"Mol. Cel. Biol."},{"key":"B51","doi-asserted-by":"publisher","first-page":"265","DOI":"10.1146\/annurev-genom-083118-014933","article-title":"The genetics and epigenetics of facioscapulohumeral muscular dystrophy","volume":"20","author":"Himeda","year":"2019","journal-title":"Annu. Rev. Genomics Hum. Genet."},{"key":"B52","doi-asserted-by":"publisher","first-page":"527","DOI":"10.1038\/mt.2015.200","article-title":"CRISPR\/dCas9-mediated transcriptional inhibition ameliorates the epigenetic dysregulation at D4Z4 and represses DUX4-fl in FSH muscular dystrophy","volume":"24","author":"Himeda","year":"2016","journal-title":"Mol. Ther."},{"key":"B53","doi-asserted-by":"publisher","first-page":"1797","DOI":"10.1016\/j.ymthe.2018.04.019","article-title":"Identification of epigenetic regulators of DUX4-fl for targeted therapy of facioscapulohumeral muscular dystrophy","volume":"26","author":"Himeda","year":"2018","journal-title":"Mol. Ther."},{"key":"B54","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1038\/nrg3863","article-title":"RNA-mediated epigenetic regulation of gene expression","volume":"16","author":"Holoch","year":"2015","journal-title":"Nat. Rev. Genet."},{"key":"B55","doi-asserted-by":"publisher","first-page":"505","DOI":"10.1146\/annurev-biophys-062215-010822","article-title":"CRISPR-Cas9 structures and mechanisms","volume":"46","author":"Jiang","year":"2017","journal-title":"Annu. Rev. Biophys."},{"key":"B56","doi-asserted-by":"publisher","first-page":"816","DOI":"10.1126\/science.1225829","article-title":"A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity","volume":"337","author":"Jinek","year":"2012","journal-title":"Science"},{"key":"B57","doi-asserted-by":"publisher","first-page":"e0192657","DOI":"10.1371\/journal.pone.0192657","article-title":"A cre-inducible DUX4 transgenic mouse model for investigating facioscapulohumeral muscular dystrophy","volume":"13","author":"Jones","year":"2018","journal-title":"PLoS One"},{"key":"B58","doi-asserted-by":"publisher","first-page":"6518","DOI":"10.1093\/nar\/gkw236","article-title":"The delivery of therapeutic oligonucleotides","volume":"44","author":"Juliano","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"B59","doi-asserted-by":"publisher","first-page":"93","DOI":"10.1089\/nat.2015.0576","article-title":"Development of a method for profiling protein interactions with LNA-modified antisense oligonucleotides using protein microarrays","volume":"26","author":"Kakiuchi-Kiyota","year":"2016","journal-title":"Nucleic Acid Ther."},{"key":"B60","doi-asserted-by":"publisher","first-page":"8638","DOI":"10.1093\/nar\/gkv857","article-title":"In silico and in vitro evaluation of exonic and intronic off-target effects form a critical element of therapeutic ASO gapmer optimization","volume":"43","author":"Kamola","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"B61","doi-asserted-by":"publisher","first-page":"237","DOI":"10.1038\/nmeth.3284","article-title":"Digenome-seq: genome-wide profiling of CRISPR-Cas9 off-target effects in human cells","volume":"12","author":"Kim","year":"2015","journal-title":"Nat. Methods"},{"key":"B62","doi-asserted-by":"publisher","first-page":"321","DOI":"10.1038\/nbt940","article-title":"Interferon induction by siRNAs and ssRNAs synthesized by phage polymerase","volume":"22","author":"Kim","year":"2004","journal-title":"Nat. Biotechnol."},{"key":"B63","doi-asserted-by":"publisher","first-page":"14500","DOI":"10.1038\/ncomms14500","article-title":"In vivo genome editing with a small Cas9 orthologue derived from Campylobacter jejuni","volume":"8","author":"Kim","year":"2017","journal-title":"Nat. Commun."},{"key":"B64","doi-asserted-by":"publisher","first-page":"367","DOI":"10.1101\/gr.231936.117","article-title":"CRISPR RNAs trigger innate immune responses in human cells","volume":"28","author":"Kim","year":"2018","journal-title":"Genome Res."},{"key":"B65","doi-asserted-by":"publisher","first-page":"553","DOI":"10.1016\/j.nmd.2006.06.008","article-title":"Severe phenotype in infantile facioscapulohumeral muscular dystrophy","volume":"16","author":"Klinge","year":"2006","journal-title":"Neuromuscul. Disord."},{"key":"B66","doi-asserted-by":"publisher","first-page":"4573","DOI":"10.1096\/fj.201902696","article-title":"DNA aptamers against the DUX4 protein reveal novel therapeutic implications for FSHD","volume":"34","author":"Klingler","year":"2020","journal-title":"FASEB J."},{"key":"B67","doi-asserted-by":"publisher","first-page":"1454","DOI":"10.1212\/wnl.51.5.1454","article-title":"Cardiac involvement in genetically confirmed facioscapulohumeral muscular dystrophy","volume":"51","author":"Lafor\u00eat","year":"1998","journal-title":"Neurology"},{"key":"B68","doi-asserted-by":"publisher","first-page":"2886","DOI":"10.3390\/jcm9092886","article-title":"Therapeutic strategies targeting DUX4 in FSHD","volume":"9","author":"Le Gall","year":"2020","journal-title":"J. Clin. Med."},{"key":"B69","doi-asserted-by":"publisher","first-page":"144","DOI":"10.3390\/jpm3030144","article-title":"Antisense therapy in neurology","volume":"3","author":"Lee","year":"2013","journal-title":"J. Pers. Med."},{"key":"B70","doi-asserted-by":"publisher","first-page":"eaay0271","DOI":"10.1126\/scitranslmed.aay0271","article-title":"Applying genome-wide CRISPR-Cas9 screens for therapeutic discovery in facioscapulohumeral muscular dystrophy","volume":"12","author":"Lek","year":"2020","journal-title":"Sci. Transl. Med."},{"key":"B71","doi-asserted-by":"publisher","first-page":"235","DOI":"10.1038\/ng999","article-title":"Facioscapulohumeral muscular dystrophy is uniquely associated with one of the two variants of the 4q subtelomere","volume":"32","author":"Lemmers","year":"2002","journal-title":"Nat. Genet."},{"key":"B72","doi-asserted-by":"publisher","first-page":"659","DOI":"10.1093\/hmg\/ddu486","article-title":"Inter-individual differences in CpG methylation at D4Z4 correlate with clinical variability in FSHD1 and FSHD2","volume":"24","author":"Lemmers","year":"2015","journal-title":"Hum. Mol. Genet."},{"key":"B73","doi-asserted-by":"publisher","first-page":"1370","DOI":"10.1038\/ng.2454","article-title":"Digenic inheritance of an SMCHD1 mutation and an FSHD-permissive D4Z4 allele causes facioscapulohumeral muscular dystrophy type 2","volume":"44","author":"Lemmers","year":"2012","journal-title":"Nat. Genet."},{"key":"B74","doi-asserted-by":"publisher","first-page":"1650","DOI":"10.1126\/science.1189044","article-title":"A unifying genetic model for facioscapulohumeral muscular dystrophy","volume":"329","author":"Lemmers","year":"2010","journal-title":"Science"},{"key":"B75","doi-asserted-by":"publisher","first-page":"1124","DOI":"10.1086\/426035","article-title":"Contractions of D4Z4 on 4qB subtelomeres do not cause facioscapulohumeral muscular dystrophy","volume":"75","author":"Lemmers","year":"2004","journal-title":"Am. J. Hum. Genet."},{"key":"B76","doi-asserted-by":"publisher","first-page":"2927","DOI":"10.1093\/nar\/gkv143","article-title":"Identification and characterization of intracellular proteins that bind oligonucleotides with phosphorothioate linkages","volume":"43","author":"Liang","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"B77","doi-asserted-by":"publisher","first-page":"4817","DOI":"10.1093\/hmg\/ddv206","article-title":"DICER\/AGO-dependent epigenetic silencing of D4Z4 repeats enhanced by exogenous siRNA suggests mechanisms and therapies for FSHD","volume":"24","author":"Lim","year":"2015","journal-title":"Hum. Mol. Genet."},{"key":"B78","doi-asserted-by":"publisher","first-page":"848","DOI":"10.1016\/j.ymthe.2020.10.010","article-title":"DUX4 transcript knockdown with antisense 2\u2032-O-methoxyethyl gapmers for the treatment of facioscapulohumeral muscular dystrophy","volume":"29","author":"Lim","year":"","journal-title":"Mol. Ther."},{"key":"B79","doi-asserted-by":"publisher","first-page":"16509","DOI":"10.1073\/pnas.1909649117","article-title":"Inhibition of DUX4 expression with antisense LNA gapmers as a therapy for facioscapulohumeral muscular dystrophy","volume":"117","author":"Lim","year":"","journal-title":"Proc. Natl. Acad. Sci. U S A."},{"key":"B80","doi-asserted-by":"publisher","first-page":"729","DOI":"10.3390\/ijms21030729","article-title":"DUX4 signalling in the pathogenesis of facioscapulohumeral muscular dystrophy","volume":"21","author":"Lim","year":"","journal-title":"Int. J. Mol. Sci."},{"key":"B81","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2018.10.011","article-title":"Efficacy of multi-exon skipping treatment in Duchenne muscular dystrophy dog model neonates","volume":"27","author":"Lim","year":"2019","journal-title":"Mol. Ther."},{"key":"B82","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1007\/978-1-4939-8651-4_1","article-title":"Invention and early history of exon skipping and splice modulation","volume-title":"Exon skipping and inclusion therapies: methods and protocols","author":"Lim","year":"2018"},{"key":"B83","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1007\/978-1-0716-0771-8_1","article-title":"Invention and early history of gapmers","volume":"2176","author":"Lim","year":"2020","journal-title":"Methods Mol. Biol"},{"key":"B84","doi-asserted-by":"publisher","first-page":"1234","DOI":"10.1080\/10717544.2018.1474964","article-title":"Delivering CRISPR: a review of the challenges and approaches","volume":"25","author":"Lino","year":"2018","journal-title":"Drug Deliv."},{"key":"B85","doi-asserted-by":"publisher","first-page":"951","DOI":"10.1093\/hmg\/4.5.951","article-title":"Correlation between fragment size at D4F104S1 and age at onset or at wheelchair use, with a possible generational effect, accounts for much phenotypic variation in 4q35-facioscapulohumeral muscular dystrophy (FSHD)","volume":"4","author":"Lunt","year":"1995","journal-title":"Hum. Mol. Genet."},{"key":"B86","doi-asserted-by":"publisher","first-page":"1374","DOI":"10.1212\/WNL.0b013e3182a84140","article-title":"Clinical and genetic features of hearing loss in facioscapulohumeral muscular dystrophy","volume":"81","author":"Lutz","year":"2013","journal-title":"Neurology"},{"key":"B87","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtn.2020.10.028","article-title":"A deoxyribonucleic acid decoy trapping DUX4 for the treatment of facioscapulohumeral muscular dystrophy","author":"Mariot","year":"2020","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"B88","doi-asserted-by":"publisher","first-page":"1468","DOI":"10.1093\/hmg\/ddw015","article-title":"Antisense targeting of 3' end elements involved in DUX4 mRNA processing is an efficient therapeutic strategy for facioscapulohumeral dystrophy: a new gene-silencing approach","volume":"25","author":"Marsollier","year":"2016","journal-title":"Hum. Mol. Genet."},{"key":"B89","doi-asserted-by":"publisher","first-page":"62","DOI":"10.1016\/j.xphs.2019.10.003","article-title":"Immunogenicity of Cas9 protein","volume":"109","author":"Mehta","year":"2020","journal-title":"J. Pharm. Sci."},{"key":"B90","doi-asserted-by":"publisher","first-page":"bio033977","DOI":"10.1242\/bio.033977","article-title":"Functional domains of the FSHD-associated DUX4 protein","volume":"7","author":"Mitsuhashi","year":"2018","journal-title":"Biol. Open"},{"key":"B91","doi-asserted-by":"publisher","first-page":"568","DOI":"10.1093\/hmg\/dds467","article-title":"Expression of DUX4 in zebrafish development recapitulates facioscapulohumeral muscular dystrophy","volume":"22","author":"Mitsuhashi","year":"2013","journal-title":"Hum. Mol. Genet."},{"key":"B92","doi-asserted-by":"publisher","first-page":"733","DOI":"10.1099\/mic.0.023960-0","article-title":"Short motif sequences determine the targets of the prokaryotic CRISPR defence system","volume":"155","author":"Mojica","year":"2009","journal-title":"Microbiology"},{"key":"B93","doi-asserted-by":"publisher","first-page":"1297","DOI":"10.1093\/nar\/gku1326","article-title":"CRISPR-based self-cleaving mechanism for controllable gene delivery in human cells","volume":"43","author":"Moore","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"B94","doi-asserted-by":"publisher","first-page":"613","DOI":"10.2144\/000113005","article-title":"Vivo-Morpholinos: a non-peptide transporter delivers Morpholinos into a wide array of mouse tissues","volume":"45","author":"Morcos","year":"2008","journal-title":"Biotechniques"},{"key":"B95","doi-asserted-by":"publisher","first-page":"1299","DOI":"10.3727\/096368910X546571","article-title":"Mesoangioblasts from facioscapulohumeral muscular dystrophy display in Vivo a variable myogenic ability predictable by their in Vitro behavior","volume":"20","author":"Morosetti","year":"2011","journal-title":"Cell Transplant."},{"key":"B96","first-page":"1202","article-title":"Antisense oligonucleotides for the treatment of cardiomyopathy in Duchenne muscular dystrophy","volume":"11","author":"Nguyen","year":"2019","journal-title":"Am. J. Transl. Res."},{"key":"B97","doi-asserted-by":"publisher","first-page":"e007798","DOI":"10.1136\/bmjopen-2015-007798","article-title":"Clinical expression of facioscapulohumeral muscular dystrophy in carriers of 1-3 D4Z4 reduced alleles: experience of the FSHD Italian national registry","volume":"6","author":"Nikolic","year":"2016","journal-title":"BMJ Open"},{"key":"B98","doi-asserted-by":"publisher","first-page":"263","DOI":"10.1038\/nature13769","article-title":"Programmable RNA recognition and cleavage by CRISPR\/Cas9","volume":"516","author":"O\u2019Connell","year":"2014","journal-title":"Nature"},{"key":"B99","doi-asserted-by":"publisher","first-page":"S73","DOI":"10.1002\/mus.880181314","article-title":"On the significance of retinal vascular disease and hearing loss in facioscapulohumeral muscular dystrophy","volume":"2","author":"Padberg","year":"1995","journal-title":"Muscle Nerve Suppl."},{"key":"B100","doi-asserted-by":"publisher","first-page":"629","DOI":"10.1242\/bio.20121305","article-title":"Conditional over-expression of PITX1 causes skeletal muscle dystrophy in mice","volume":"1","author":"Pandey","year":"2012","journal-title":"Biol. Open"},{"key":"B101","doi-asserted-by":"publisher","first-page":"8304","DOI":"10.3390\/molecules20058304","article-title":"Culture conditions affect expression of DUX4 in FSHD myoblasts","volume":"20","author":"Pandey","year":"2015","journal-title":"Molecules"},{"key":"B102","doi-asserted-by":"publisher","first-page":"390","DOI":"10.1038\/mt.2013.263","article-title":"Morpholino treatment improves muscle function and pathology of Pitx1 transgenic mice","volume":"22","author":"Pandey","year":"2014","journal-title":"Mol. Ther."},{"key":"B103","doi-asserted-by":"publisher","first-page":"1380","DOI":"10.1016\/j.cell.2013.08.021","article-title":"Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity","volume":"154","author":"Ran","year":"2013","journal-title":"Cell"},{"key":"B104","doi-asserted-by":"publisher","first-page":"3408","DOI":"10.1093\/brain\/awt226","article-title":"Large scale genotype-phenotype analyses indicate that novel prognostic tools are required for families with facioscapulohumeral muscular dystrophy","volume":"136","author":"Ricci","year":"2013","journal-title":"Brain"},{"key":"B105","doi-asserted-by":"publisher","first-page":"325","DOI":"10.1007\/s00439-011-1100-z","article-title":"Facioscapulohumeral muscular dystrophy (FSHD): an enigma unravelled?","volume":"131","author":"Richards","year":"2012","journal-title":"Hum. Genet."},{"key":"B106","doi-asserted-by":"publisher","first-page":"5901","DOI":"10.1093\/hmg\/ddv315","article-title":"Endogenous DUX4 expression in FSHD myotubes is sufficient to cause cell death and disrupts RNA splicing and cell migration pathways","volume":"24","author":"Rickard","year":"2015","journal-title":"Hum. Mol. Genet."},{"key":"B107","doi-asserted-by":"publisher","first-page":"3643","DOI":"10.3390\/ijms21103643","article-title":"High-capacity adenoviral vectors: expanding the scope of gene therapy","volume":"21","author":"Ricobaraza","year":"2020","journal-title":"Int. J. Mol. Sci."},{"key":"B108","doi-asserted-by":"publisher","first-page":"283","DOI":"10.1002\/mus.21244","article-title":"Inhibition of myostatin with emphasis on follistatin as a therapy for muscle disease","volume":"39","author":"Rodino-Klapac","year":"2009","journal-title":"Muscle Nerve"},{"key":"B109","doi-asserted-by":"publisher","first-page":"e2273","DOI":"10.1212\/WNL.0000000000007456","article-title":"FSHD1 and FSHD2 form a disease continuum","volume":"92","author":"Sacconi","year":"2019","journal-title":"Neurology"},{"key":"B110","doi-asserted-by":"publisher","first-page":"744","DOI":"10.1016\/j.ajhg.2013.08.004","article-title":"The FSHD2 gene SMCHD1 is a modifier of disease severity in families affected by FSHD1","volume":"93","author":"Sacconi","year":"2013","journal-title":"Am. J. Hum. Genet."},{"key":"B111","doi-asserted-by":"publisher","first-page":"739","DOI":"10.1002\/mus.24218","article-title":"Restrictive lung involvement in facioscapulohumeral muscular dystrophy","volume":"50","author":"Scully","year":"2014","journal-title":"Muscle Nerve"},{"key":"B112","doi-asserted-by":"publisher","first-page":"399","DOI":"10.1038\/nmeth.2857","article-title":"Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects","volume":"11","author":"Shen","year":"2014","journal-title":"Nat. Methods"},{"key":"B113","doi-asserted-by":"publisher","first-page":"2204","DOI":"10.1093\/nar\/gky060","article-title":"Acute hepatotoxicity of 2' fluoro-modified 5-10-5 gapmer phosphorothioate oligonucleotides in mice correlates with intracellular protein binding and the loss of DBHS proteins","volume":"46","author":"Shen","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"B114","doi-asserted-by":"publisher","first-page":"640","DOI":"10.1038\/s41587-019-0106-2","article-title":"Chemical modification of PS-ASO therapeutics reduces cellular protein-binding and improves the therapeutic index","volume":"37","author":"Shen","year":"2019","journal-title":"Nat. Biotechnol."},{"key":"B115","doi-asserted-by":"publisher","first-page":"709","DOI":"10.1016\/j.ymthe.2020.01.001","article-title":"Immune responses to viral gene therapy vectors","volume":"28","author":"Shirley","year":"2020","journal-title":"Mol. Ther."},{"key":"B116","doi-asserted-by":"publisher","first-page":"e1001181","DOI":"10.1371\/journal.pgen.1001181","article-title":"Facioscapulohumeral dystrophy: incomplete suppression of a retrotransposed gene","volume":"6","author":"Snider","year":"2010","journal-title":"Plos Genet."},{"key":"B117","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1089\/oli.1.1997.7.187","article-title":"Morpholino antisense oligomers: design, preparation, and properties","volume":"7","author":"Summerton","year":"1997","journal-title":"Antisense Nucleic Acid Drug Dev."},{"key":"B118","doi-asserted-by":"publisher","first-page":"76","DOI":"10.1111\/j.1582-4934.2012.01647.x","article-title":"DUX4 expression in FSHD muscle cells: how could such a rare protein cause a myopathy?","volume":"17","author":"Tassin","year":"2013","journal-title":"J. Cel. Mol. Med."},{"key":"B119","doi-asserted-by":"publisher","first-page":"12","DOI":"10.1186\/2044-5040-4-12","article-title":"Facioscapulohumeral dystrophy: the path to consensus on pathophysiology","volume":"4","author":"Tawil","year":"2014","journal-title":"Skelet. Muscle"},{"key":"B120","doi-asserted-by":"publisher","first-page":"726","DOI":"10.1038\/s41578-019-0145-9","article-title":"Engineered materials for in vivo delivery of genome-editing machinery","volume":"4","author":"Tong","year":"2019","journal-title":"Nat. Rev. Mater."},{"key":"B121","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1038\/nbt.3117","article-title":"GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases","volume":"33","author":"Tsai","year":"2015","journal-title":"Nat. Biotechnol."},{"key":"B122","doi-asserted-by":"publisher","first-page":"355","DOI":"10.1016\/j.molcel.2016.07.004","article-title":"Methods for optimizing CRISPR-cas9 genome editing specificity","volume":"63","author":"Tycko","year":"2016","journal-title":"Mol. Cell"},{"key":"B123","doi-asserted-by":"publisher","first-page":"1020","DOI":"10.1016\/j.ajhg.2016.03.013","article-title":"Mutations in DNMT3B modify epigenetic repression of the D4Z4 repeat and the penetrance of facioscapulohumeral dystrophy","volume":"98","author":"van den Boogaard","year":"2016","journal-title":"Am. J. Hum. Genet."},{"key":"B124","doi-asserted-by":"publisher","first-page":"569","DOI":"10.1002\/ana.20625","article-title":"Variable hypomethylation of D4Z4 in facioscapulohumeral muscular dystrophy","volume":"58","author":"Van Overveld","year":"2005","journal-title":"Ann. Neurol."},{"key":"B125","doi-asserted-by":"publisher","first-page":"315","DOI":"10.1038\/ng1262","article-title":"Hypomethylation of D4Z4 in 4q-linked and non-4q-linked facioscapulohumeral muscular dystrophy","volume":"35","author":"van Overveld","year":"2003","journal-title":"Nat. Genet."},{"key":"B126","doi-asserted-by":"publisher","first-page":"e26820","DOI":"10.1371\/journal.pone.0026820","article-title":"The FSHD atrophic myotube phenotype is caused by DUX4 expression","volume":"6","author":"Vanderplanck","year":"2011","journal-title":"PLoS One"},{"key":"B127","doi-asserted-by":"publisher","first-page":"1651","DOI":"10.1038\/sj.gt.3302565","article-title":"Normal growth and regenerating ability of myoblasts from unaffected muscles of facioscapulohumeral muscular dystrophy patients","volume":"12","author":"Vilquin","year":"2005","journal-title":"Gene Ther."},{"key":"B128","doi-asserted-by":"publisher","first-page":"242","DOI":"10.1038\/s41591-018-0204-6","article-title":"High prevalence of Streptococcus pyogenes Cas9-reactive T cells within the adult human population","volume":"25","author":"Wagner","year":"2019","journal-title":"Nat. Med."},{"key":"B129","doi-asserted-by":"publisher","first-page":"1417","DOI":"10.1038\/mt.2012.68","article-title":"RNA interference inhibits DUX4-induced muscle toxicity in vivo: implications for a targeted FSHD therapy","volume":"20","author":"Wallace","year":"2012","journal-title":"Mol. Ther."},{"key":"B130","doi-asserted-by":"publisher","first-page":"121","DOI":"10.1016\/j.omtm.2017.12.005","article-title":"Pre-clinical safety and off-target studies to support translation of AAV-mediated RNAi therapy for FSHD","volume":"8","author":"Wallace","year":"2018","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"B131","doi-asserted-by":"publisher","first-page":"4284","DOI":"10.1038\/s41467-019-12281-8","article-title":"Optimized CRISPR guide RNA design for two high-fidelity Cas9 variants by deep learning","volume":"10","author":"Wang","year":"2019","journal-title":"Nat. Commun."},{"key":"B132","doi-asserted-by":"publisher","first-page":"66","DOI":"10.1007\/s11910-016-0667-0","article-title":"Facioscapulohumeral dystrophy","volume":"16","author":"Wang","year":"2016","journal-title":"Curr. Neurol. Neurosci. Rep."},{"key":"B133","doi-asserted-by":"publisher","first-page":"e2005840","DOI":"10.1371\/journal.pbio.2005840","article-title":"In vitro-transcribed guide RNAs trigger an innate immune response via the RIG-I pathway","volume":"16","author":"Wienert","year":"2018","journal-title":"PLoS Biol."},{"key":"B134","doi-asserted-by":"publisher","first-page":"1555","DOI":"10.1021\/acs.accounts.9b00106","article-title":"Delivery aspects of CRISPR\/Cas for in Vivo genome editing","volume":"52","author":"Wilbie","year":"2019","journal-title":"Acc. Chem. Res."},{"key":"B135","doi-asserted-by":"publisher","first-page":"80","DOI":"10.1038\/mt.2009.255","article-title":"Effect of genome size on AAV vector packaging","volume":"18","author":"Wu","year":"2010","journal-title":"Mol. Ther."},{"key":"B136","doi-asserted-by":"publisher","first-page":"28","DOI":"10.3390\/v11010028","article-title":"Viral delivery systems for CRISPR","volume":"11","author":"Xu","year":"2019","journal-title":"Viruses"},{"key":"B137","doi-asserted-by":"publisher","first-page":"827","DOI":"10.1111\/gtc.12730","article-title":"Evaluation of off-target effects of gapmer antisense oligonucleotides using human cells","volume":"24","author":"Yoshida","year":"2019","journal-title":"Genes Cells"}],"container-title":["Frontiers in Pharmacology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2021.642858\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,3,12]],"date-time":"2021-03-12T16:09:49Z","timestamp":1615565389000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2021.642858\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,3,12]]},"references-count":137,"alternative-id":["10.3389\/fphar.2021.642858"],"URL":"https:\/\/doi.org\/10.3389\/fphar.2021.642858","relation":{},"ISSN":["1663-9812"],"issn-type":[{"value":"1663-9812","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,3,12]]},"article-number":"642858"}}