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Characterisation of Non-Pathogenic Premutation-Range Myotonic Dystrophy Type 2 Alleles
Article

Characterization of RAN Translation and Antisense Transcription in Primary Cell Cultures of Patients with Myotonic Dystrophy Type 1

1
Neuromuscular and Neuropediatric Research Group, Institut d’Investigació en Ciències de la Salut Germans Trias i Pujol (IGTP), Campus Can Ruti, Universitat Autònoma de Barcelona, 08916 Badalona, Spain
2
Centre for Biomedical Network Research on Rare Diseases (CIBERER), Instituto de Salud Carlos III, 28029 Madrid, Spain
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Neuromuscular Pathology Unit, Neurology Service, Neuroscience Department, Hospital Universitari Germans Trias i Pujol, 08916 Badalona, Spain
4
IrsiCaixa AIDS Research Institute, 08916 Badalona, Spain
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Laboratory of Molecular, Cellular and Genomic Biomedicine, Instituto de Investigación Sanitaria La Fe, 46026 Valencia, Spain
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Joint Unit for Rare Diseases IIS La Fe-CIPF, 46012 Valencia, Spain
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Reference Unit for Hereditary Metabolic Disorders (MetabERN), Vall d’Hebron University Hospital, 08035 Barcelona, Spain
*
Author to whom correspondence should be addressed.
Equal contribution.
Academic Editor: Sabrina Ravaglia
J. Clin. Med. 2021, 10(23), 5520; https://doi.org/10.3390/jcm10235520 (registering DOI)
Received: 30 October 2021 / Revised: 18 November 2021 / Accepted: 19 November 2021 / Published: 25 November 2021
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. View Full-Text
Keywords: RAN translation; antisense transcription; myotonic dystrophies; primary cell cultures; phenotypic modulators RAN translation; antisense transcription; myotonic dystrophies; primary cell cultures; phenotypic modulators
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MDPI and ACS Style

Koehorst, E.; Núñez-Manchón, J.; Ballester-López, A.; Almendrote, M.; Lucente, G.; Arbex, A.; Chojnacki, J.; Vázquez-Manrique, R.P.; Gómez-Escribano, A.P.; Pintos-Morell, G.; Coll-Cantí, J.; Ramos-Fransi, A.; Martínez-Piñeiro, A.; Suelves, M.; Nogales-Gadea, G. Characterization of RAN Translation and Antisense Transcription in Primary Cell Cultures of Patients with Myotonic Dystrophy Type 1. J. Clin. Med. 2021, 10, 5520. https://doi.org/10.3390/jcm10235520

AMA Style

Koehorst E, Núñez-Manchón J, Ballester-López A, Almendrote M, Lucente G, Arbex A, Chojnacki J, Vázquez-Manrique RP, Gómez-Escribano AP, Pintos-Morell G, Coll-Cantí J, Ramos-Fransi A, Martínez-Piñeiro A, Suelves M, Nogales-Gadea G. Characterization of RAN Translation and Antisense Transcription in Primary Cell Cultures of Patients with Myotonic Dystrophy Type 1. Journal of Clinical Medicine. 2021; 10(23):5520. https://doi.org/10.3390/jcm10235520

Chicago/Turabian Style

Koehorst, Emma, Judit Núñez-Manchón, Alfonsina Ballester-López, Miriam Almendrote, Giuseppe Lucente, Andrea Arbex, Jakub Chojnacki, Rafael P. Vázquez-Manrique, Ana P. Gómez-Escribano, Guillem Pintos-Morell, Jaume Coll-Cantí, Alba Ramos-Fransi, Alicia Martínez-Piñeiro, Mònica Suelves, and Gisela Nogales-Gadea. 2021. "Characterization of RAN Translation and Antisense Transcription in Primary Cell Cultures of Patients with Myotonic Dystrophy Type 1" Journal of Clinical Medicine 10, no. 23: 5520. https://doi.org/10.3390/jcm10235520

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