{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T13:27:19Z","timestamp":1790774839499,"version":"4.1.0"},"reference-count":29,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2021,11,25]],"date-time":"2021-11-25T00:00:00Z","timestamp":1637798400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["PI15\/01756, PI18\/00713, IFI20\/00022, CPII16\/00004, PI17\/00011 and PI20\/00114"],"award-info":[{"award-number":["PI15\/01756, PI18\/00713, IFI20\/00022, CPII16\/00004, PI17\/00011 and PI20\/00114"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007393","name":"French Muscular Dystrophy Association","doi-asserted-by":"publisher","award":["#21108"],"award-info":[{"award-number":["#21108"]}],"id":[{"id":"10.13039\/100007393","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Miquel Servet grant","award":["ISCIII CD14\/00032, ISCIII CPII19\/00021"],"award-info":[{"award-number":["ISCIII CD14\/00032, ISCIII CPII19\/00021"]}]},{"DOI":"10.13039\/501100004837","name":"Ministerio de Ciencia e Innovaci\u00f3n","doi-asserted-by":"publisher","award":["PID2020-118730RB-I00"],"award-info":[{"award-number":["PID2020-118730RB-I00"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"name":"European Union's Horizon 2020 grant","award":["793830"],"award-info":[{"award-number":["793830"]}]},{"DOI":"10.13039\/100008054","name":"Fundaci\u00f3n Ram\u00f3n Areces","doi-asserted-by":"publisher","award":["CIVP19S8119"],"award-info":[{"award-number":["CIVP19S8119"]}],"id":[{"id":"10.13039\/100008054","id-type":"DOI","asserted-by":"publisher"}]},{"name":"la Caixa Foundation INPhINIT fellowship","award":["LCF\/BQ\/IN18\/11660019"],"award-info":[{"award-number":["LCF\/BQ\/IN18\/11660019"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JCM"],"abstract":"<jats:p>Myotonic Dystrophy type 1 (DM1) is a muscular dystrophy with a multi-systemic nature. It was one of the first diseases in which repeat associated non-ATG (RAN) translation was described in 2011, but has not been further explored since. In order to enhance our knowledge of RAN translation in DM1, we decided to study the presence of DM1 antisense (DM1-AS) transcripts (the origin of the polyglutamine (polyGln) RAN protein) using RT-PCR and FISH, and that of RAN translation via immunoblotting and immunofluorescence in distinct DM1 primary cell cultures, e.g., myoblasts, skin fibroblasts and lymphoblastoids, from ten patients. DM1-AS transcripts were found in all DM1 cells, with a lower expression in patients compared to controls. Antisense RNA foci were found in the nuclei and cytoplasm of a subset of DM1 cells. The polyGln RAN protein was undetectable in all three cell types with both approaches. Immunoblots revealed a 42 kD polyGln containing protein, which was most likely the TATA-box-binding protein. Immunofluorescence revealed a cytoplasmic aggregate, which co-localized with the Golgi apparatus. Taken together, DM1-AS transcript levels were lower in patients compared to controls and a small portion of the transcripts included the expanded repeat. However, RAN translation was not present in patient-derived DM1 cells, or was in undetectable quantities for the available methods.<\/jats:p>","DOI":"10.3390\/jcm10235520","type":"journal-article","created":{"date-parts":[[2021,12,1]],"date-time":"2021-12-01T01:43:22Z","timestamp":1638323002000},"page":"5520","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Characterization of RAN Translation and Antisense Transcription in Primary Cell Cultures of Patients with Myotonic Dystrophy Type 1"],"prefix":"10.3390","volume":"10","author":[{"given":"Emma","family":"Koehorst","sequence":"first","affiliation":[{"name":"Neuromuscular and Neuropediatric Research Group, Institut d\u2019Investigaci\u00f3 en Ci\u00e8ncies de la Salut Germans Trias i Pujol (IGTP), Campus Can Ruti, Universitat Aut\u00f2noma de Barcelona, 08916 Badalona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Judit","family":"N\u00fa\u00f1ez-Manch\u00f3n","sequence":"additional","affiliation":[{"name":"Neuromuscular and Neuropediatric Research Group, Institut d\u2019Investigaci\u00f3 en Ci\u00e8ncies de la Salut Germans Trias i Pujol (IGTP), Campus Can Ruti, Universitat Aut\u00f2noma de Barcelona, 08916 Badalona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alfonsina","family":"Ballester-L\u00f3pez","sequence":"additional","affiliation":[{"name":"Neuromuscular and Neuropediatric Research Group, Institut d\u2019Investigaci\u00f3 en Ci\u00e8ncies de la Salut Germans Trias i Pujol (IGTP), Campus Can Ruti, Universitat Aut\u00f2noma de Barcelona, 08916 Badalona, Spain"},{"name":"Centre for Biomedical Network Research on Rare Diseases (CIBERER), Instituto de Salud Carlos III, 28029 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miriam","family":"Almendrote","sequence":"additional","affiliation":[{"name":"Neuromuscular and Neuropediatric Research Group, Institut d\u2019Investigaci\u00f3 en Ci\u00e8ncies de la Salut Germans Trias i Pujol (IGTP), Campus Can Ruti, Universitat Aut\u00f2noma de Barcelona, 08916 Badalona, Spain"},{"name":"Neuromuscular Pathology Unit, Neurology Service, Neuroscience Department, Hospital Universitari Germans Trias i Pujol, 08916 Badalona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1120-9136","authenticated-orcid":false,"given":"Giuseppe","family":"Lucente","sequence":"additional","affiliation":[{"name":"Neuromuscular and Neuropediatric Research Group, Institut d\u2019Investigaci\u00f3 en Ci\u00e8ncies de la Salut Germans Trias i Pujol (IGTP), Campus Can Ruti, Universitat Aut\u00f2noma de Barcelona, 08916 Badalona, Spain"},{"name":"Neuromuscular Pathology Unit, Neurology Service, Neuroscience Department, Hospital Universitari Germans Trias i Pujol, 08916 Badalona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea","family":"Arbex","sequence":"additional","affiliation":[{"name":"Neuromuscular and Neuropediatric Research Group, Institut d\u2019Investigaci\u00f3 en Ci\u00e8ncies de la Salut Germans Trias i Pujol (IGTP), Campus Can Ruti, Universitat Aut\u00f2noma de Barcelona, 08916 Badalona, Spain"},{"name":"Neuromuscular Pathology Unit, Neurology Service, Neuroscience Department, Hospital Universitari Germans Trias i Pujol, 08916 Badalona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jakub","family":"Chojnacki","sequence":"additional","affiliation":[{"name":"IrsiCaixa AIDS Research Institute, 08916 Badalona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1594-0033","authenticated-orcid":false,"given":"Rafael","family":"V\u00e1zquez-Manrique","sequence":"additional","affiliation":[{"name":"Centre for Biomedical Network Research on Rare Diseases (CIBERER), Instituto de Salud Carlos III, 28029 Madrid, Spain"},{"name":"Laboratory of Molecular, Cellular and Genomic Biomedicine, Instituto de Investigaci\u00f3n Sanitaria La Fe, 46026 Valencia, Spain"},{"name":"Joint Unit for Rare Diseases IIS La Fe-CIPF, 46012 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4391-5932","authenticated-orcid":false,"given":"Ana","family":"G\u00f3mez-Escribano","sequence":"additional","affiliation":[{"name":"Centre for Biomedical Network Research on Rare Diseases (CIBERER), Instituto de Salud Carlos III, 28029 Madrid, Spain"},{"name":"Laboratory of Molecular, Cellular and Genomic Biomedicine, Instituto de Investigaci\u00f3n Sanitaria La Fe, 46026 Valencia, Spain"},{"name":"Joint Unit for Rare Diseases IIS La Fe-CIPF, 46012 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guillem","family":"Pintos-Morell","sequence":"additional","affiliation":[{"name":"Neuromuscular and Neuropediatric Research Group, Institut d\u2019Investigaci\u00f3 en Ci\u00e8ncies de la Salut Germans Trias i Pujol (IGTP), Campus Can Ruti, Universitat Aut\u00f2noma de Barcelona, 08916 Badalona, Spain"},{"name":"Reference Unit for Hereditary Metabolic Disorders (MetabERN), Vall d\u2019Hebron University Hospital, 08035 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jaume","family":"Coll-Cant\u00ed","sequence":"additional","affiliation":[{"name":"Neuromuscular and Neuropediatric Research Group, Institut d\u2019Investigaci\u00f3 en Ci\u00e8ncies de la Salut Germans Trias i Pujol (IGTP), Campus Can Ruti, Universitat Aut\u00f2noma de Barcelona, 08916 Badalona, Spain"},{"name":"Neuromuscular Pathology Unit, Neurology Service, Neuroscience Department, Hospital Universitari Germans Trias i Pujol, 08916 Badalona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alba","family":"Ramos-Fransi","sequence":"additional","affiliation":[{"name":"Neuromuscular and Neuropediatric Research Group, Institut d\u2019Investigaci\u00f3 en Ci\u00e8ncies de la Salut Germans Trias i Pujol (IGTP), Campus Can Ruti, Universitat Aut\u00f2noma de Barcelona, 08916 Badalona, Spain"},{"name":"Neuromuscular Pathology Unit, Neurology Service, Neuroscience Department, Hospital Universitari Germans Trias i Pujol, 08916 Badalona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alicia","family":"Mart\u00ednez-Pi\u00f1eiro","sequence":"additional","affiliation":[{"name":"Neuromuscular and Neuropediatric Research Group, Institut d\u2019Investigaci\u00f3 en Ci\u00e8ncies de la Salut Germans Trias i Pujol (IGTP), Campus Can Ruti, Universitat Aut\u00f2noma de Barcelona, 08916 Badalona, Spain"},{"name":"Neuromuscular Pathology Unit, Neurology Service, Neuroscience Department, Hospital Universitari Germans Trias i Pujol, 08916 Badalona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M\u00f2nica","family":"Suelves","sequence":"additional","affiliation":[{"name":"Neuromuscular and Neuropediatric Research Group, Institut d\u2019Investigaci\u00f3 en Ci\u00e8ncies de la Salut Germans Trias i Pujol (IGTP), Campus Can Ruti, Universitat Aut\u00f2noma de Barcelona, 08916 Badalona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gisela","family":"Nogales-Gadea","sequence":"additional","affiliation":[{"name":"Neuromuscular and Neuropediatric Research Group, Institut d\u2019Investigaci\u00f3 en Ci\u00e8ncies de la Salut Germans Trias i Pujol (IGTP), Campus Can Ruti, Universitat Aut\u00f2noma de Barcelona, 08916 Badalona, Spain"},{"name":"Centre for Biomedical Network Research on Rare Diseases (CIBERER), Instituto de Salud Carlos III, 28029 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,11,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"594","DOI":"10.1016\/j.bbadis.2014.05.019","article-title":"Myotonic dystrophies: An update on clinical aspects, genetic, pathology, and molecular pathomechanisms","volume":"1852","author":"Meola","year":"2015","journal-title":"Biochim. Biophys. Acta Mol. Basis Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1016\/j.molcel.2005.09.002","article-title":"Antisense transcription and heterochromatin at the DM1 CTG repeats are constrained by CTCF","volume":"20","author":"Cho","year":"2005","journal-title":"Mol. Cell"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Huguet, A., Medja, F., Nicole, A., Vignaud, A., Guiraud-Dogan, C., Ferry, A., Decostre, V., Hogrel, J.Y., Metzger, F., and Hoeflich, A. (2012). Molecular, Physiological, and Motor Performance Defects in DMSXL Mice Carrying >1000 CTG Repeats from the Human DM1 Locus. PLoS Genet., 8.","DOI":"10.1371\/journal.pgen.1003043"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Michel, L., Huguet-Lachon, A., and Gourdon, G. (2015). Sense and antisense DMPK RNA foci accumulate in DM1 tissues during development. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0137620"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1374","DOI":"10.1080\/15476286.2017.1279787","article-title":"Antisense transcription of the myotonic dystrophy locus yields low-abundant RNAs with and without (CAG)n repeat","volume":"14","author":"Gudde","year":"2017","journal-title":"RNA Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1073\/pnas.1013343108","article-title":"Non-ATG-initiated translation directed by microsatellite expansions","volume":"108","author":"Zu","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1146\/annurev-neuro-070918-050405","article-title":"Repeat-Associated Non-ATG Translation: Molecular Mechanisms and Contribution to Neurological Disease","volume":"42","author":"Nguyen","year":"2019","journal-title":"Annu. Rev. Neurosci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1215","DOI":"10.1093\/nar\/16.3.1215","article-title":"A simple salting out procedure for extracting DNA from human nucleated cells","volume":"16","author":"Miller","year":"1988","journal-title":"Nucleic Acids Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1385\/1-59259-804-8:061","article-title":"Analysis of Unstable Triplet Repeats Using Small-Pool Polymerase Chain Reaction","volume":"Volume 277","author":"Bidichandani","year":"2004","journal-title":"Trinucleotide Repeat Protocols"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1093\/hmg\/4.1.1","article-title":"Somatic mosaicism, germline expansions, germline reversions and intergenerational reductions in myotonic dystrophy males: Small pool PCR analyses","volume":"4","author":"Monckton","year":"1995","journal-title":"Hum. Mol. Genet."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"pdb.prot5441","DOI":"10.1101\/pdb.prot5441","article-title":"Preparation of cytoplasmic and nuclear RNA from tissue culture cells","volume":"2010","author":"Rio","year":"2010","journal-title":"Cold Spring Harb. Protoc."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Ballester-Lopez, A., Koehorst, E., Linares-Pardo, I., N\u00fa\u00f1ez-Manch\u00f3n, J., Almendrote, M., Lucente, G., Arbex, A., Alonso, C.P., Lucia, A., and Monckton, D.G. (2020). Preliminary findings on ctg expansion determination in different tissues from patients with myotonic dystrophy type 1. Genes, 11.","DOI":"10.3390\/genes11111321"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e484","DOI":"10.1212\/NXG.0000000000000484","article-title":"Three-dimensional imaging in myotonic dystrophy type 1","volume":"6","author":"Koehorst","year":"2020","journal-title":"Neurol. Genet."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1002\/humu.23932","article-title":"A DM1 family with interruptions associated with atypical symptoms and late onset but not with a milder phenotype","volume":"41","author":"Koehorst","year":"2020","journal-title":"Hum. Mutat."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"567435","DOI":"10.1155\/2013\/567435","article-title":"Transcriptionally repressive chromatin remodelling and CpG methylation in the presence of expanded CTG-repeats at the DM1 locus","volume":"2013","author":"Brouwer","year":"2013","journal-title":"J. Nucleic Acids"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1038\/378403a0","article-title":"Polyglutamine expansion as a pathological epitope in Huntington\u2019s disease and four dominant cerebellar ataxias","volume":"378","author":"Trottier","year":"1995","journal-title":"Nature"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3558","DOI":"10.1093\/hmg\/dds185","article-title":"Somatic instability of the expanded CTG triplet repeat in myotonic dystrophy type 1 is a heritable quantitative trait and modifier of disease severity","volume":"21","author":"Morales","year":"2012","journal-title":"Hum. Mol. Genet."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2496","DOI":"10.1093\/hmg\/ddaa123","article-title":"Longitudinal increases in somatic mosaicism of the expanded CTG repeat in myotonic dystrophy type 1 are associated with variation in age-at-onset","volume":"29","author":"Morales","year":"2020","journal-title":"Hum. Mol. Genet."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1485","DOI":"10.1093\/hmg\/ddg175","article-title":"Expansion of the polyQ repeat in ataxin-2 alters its Golgi localization, disrupts the Golgi complex and causes cell death","volume":"12","author":"Huynh","year":"2003","journal-title":"Hum. Mol. Genet."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"639","DOI":"10.1016\/j.neuron.2013.02.004","article-title":"Unconventional translation of C9ORF72 GGGGCC expansion generates insoluble polypeptides specific to c9FTD\/ALS","volume":"77","author":"Ash","year":"2013","journal-title":"Neuron"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"E4968","DOI":"10.1073\/pnas.1315438110","article-title":"RAN proteins and RNA foci from antisense transcripts in C9ORF72 ALS and frontotemporal dementia","volume":"110","author":"Zu","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1335","DOI":"10.1126\/science.1232927","article-title":"The C9orf72 GGGGCC repeat is translated into aggregating dipeptide-repeat proteins in FTLD\/ALS","volume":"339","author":"Mori","year":"2013","journal-title":"Science"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1016\/j.neuron.2013.03.026","article-title":"CGG repeat-associated translation mediates neurodegeneration in fragile X tremor ataxia syndrome","volume":"78","author":"Todd","year":"2013","journal-title":"Neuron"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1016\/j.neuron.2015.10.038","article-title":"RAN Translation in Huntington Disease","volume":"88","author":"Ayhan","year":"2015","journal-title":"Neuron"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"893","DOI":"10.1136\/jmg.2006.044222","article-title":"Polyalanine and polyserine frameshift products in Huntington\u2019s disease","volume":"43","author":"Davies","year":"2006","journal-title":"J. Med. Genet."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1016\/j.neuron.2017.02.046","article-title":"Regulatory Role of RNA Chaperone TDP-43 for RNA Misfolding and Repeat-Associated Translation in SCA31","volume":"94","author":"Ishiguro","year":"2017","journal-title":"Neuron"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2649","DOI":"10.1093\/hmg\/ddi299","article-title":"Ribosomal frameshifting on MJD-1 transcripts with long CAG tracts","volume":"14","author":"Toulouse","year":"2005","journal-title":"Hum. Mol. Genet."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1888","DOI":"10.1167\/iovs.17-23265","article-title":"Repeat-Associated Non-ATG (RAN) Translation in Fuchs\u2019 Endothelial Corneal Dystrophy","volume":"59","author":"Soragni","year":"2018","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1292","DOI":"10.1016\/j.neuron.2017.08.039","article-title":"RAN Translation Regulated by Muscleblind Proteins in Myotonic Dystrophy Type 2","volume":"95","author":"Zu","year":"2017","journal-title":"Neuron"}],"container-title":["Journal of Clinical Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2077-0383\/10\/23\/5520\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:35:45Z","timestamp":1760168145000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2077-0383\/10\/23\/5520"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,11,25]]},"references-count":29,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["jcm10235520"],"URL":"https:\/\/doi.org\/10.3390\/jcm10235520","relation":{},"ISSN":["2077-0383"],"issn-type":[{"value":"2077-0383","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,11,25]]}}}