{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T06:26:24Z","timestamp":1790490384764,"version":"4.1.0"},"reference-count":43,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2020,5,18]],"date-time":"2020-05-18T00:00:00Z","timestamp":1589760000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["19K20673, 18K07845 and 18K07861"],"award-info":[{"award-number":["19K20673, 18K07845 and 18K07861"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The DMD gene is one of the largest human genes, being composed of 79 exons, and encodes dystrophin Dp427m which is deficient in Duchenne muscular dystrophy (DMD). In some DMD patient, however, small size dystrophin reacting with antibody to N-terminal but not to C-terminal has been identified. The mechanism to produce N-terminal small size dystrophin remains unknown. Intronic polyadenylation is a mechanism that produces a transcript with a new 3\u2032 terminal exon and a C-terminal truncated protein. In this study, intronic alternative polyadenylation was disclosed to occur in the middle of the DMD gene and produce the half-size N-terminal dystrophin Dp427m, Dpm234. The 3\u2032-rapid amplification of cDNA ends revealed 421 bp sequence in the downstream of DMD exon 41 in U-251 glioblastoma cells. The cloned sequence composing of the 5\u2032 end sequence of intron 41 was decided as the terminal exon, since it encoded poly (A) signal followed by poly (A) stretch. Subsequently, a fragment from DMD exon M1 to intron 41 was obtained by PCR amplification. This product was named Dpm234 after its molecular weight. However, Dpm234 was not PCR amplified in human skeletal and cardiac muscles. Remarkably, Dpm234 was PCR amplified in iPS-derived cardiomyocytes. Accordingly, Western blotting of cardiomyocyte proteins showed a band of 234 kDa reacting with dystrophin antibody to N-terminal, but not C-terminal. Clinically, DMD patients with mutations in the Dpm234 coding region were found to have a significantly higher likelihood of two ECG abnormal findings. Intronic alternative splicing was first revealed in Dp427m to produce small size dystrophin.<\/jats:p>","DOI":"10.3390\/ijms21103555","type":"journal-article","created":{"date-parts":[[2020,5,18]],"date-time":"2020-05-18T11:34:14Z","timestamp":1589801654000},"page":"3555","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Intronic Alternative Polyadenylation in the Middle of the DMD Gene Produces Half-Size N-Terminal Dystrophin with a Potential Implication of ECG Abnormalities of DMD Patients"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0809-303X","authenticated-orcid":false,"given":"Abdul Qawee Mahyoob","family":"Rani","sequence":"first","affiliation":[{"name":"Research Center for Locomotion Biology, Kobe Gakuin University, Nishi, Kobe 6512180, Japan"},{"name":"KNC Department of Nucleic Acid Drug Discovery, Faculty of Rehabilitation, Kobe Gakuin University, Nishi, Kobe 6512180, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1671-357X","authenticated-orcid":false,"given":"Tetsushi","family":"Yamamoto","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory, Kobe University Hospital, Chuo, Kobe 6500017, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tatsuya","family":"Kawaguchi","sequence":"additional","affiliation":[{"name":"Translational Research Department, Daiichi Sankyo RD Novare Co., Ltd., Edogawa, Tokyo 1348630, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kazuhiro","family":"Maeta","sequence":"additional","affiliation":[{"name":"Research Center for Locomotion Biology, Kobe Gakuin University, Nishi, Kobe 6512180, Japan"},{"name":"KNC Department of Nucleic Acid Drug Discovery, Faculty of Rehabilitation, Kobe Gakuin University, Nishi, Kobe 6512180, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9846-4142","authenticated-orcid":false,"given":"Hiroyuki","family":"Awano","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Kobe University Graduate School of Medicine, Chuo, Kobe 6500017, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6778-4511","authenticated-orcid":false,"given":"Hisahide","family":"Nishio","sequence":"additional","affiliation":[{"name":"Research Center for Locomotion Biology, Kobe Gakuin University, Nishi, Kobe 6512180, Japan"},{"name":"Department of Occupational Therapy, Faculty of Rehabilitation, Kobe Gakuin University, Nishi, Kobe 6512180, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6747-7194","authenticated-orcid":false,"given":"Masafumi","family":"Matsuo","sequence":"additional","affiliation":[{"name":"Research Center for Locomotion Biology, Kobe Gakuin University, Nishi, Kobe 6512180, Japan"},{"name":"KNC Department of Nucleic Acid Drug Discovery, Faculty of Rehabilitation, Kobe Gakuin University, Nishi, Kobe 6512180, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,5,18]]},"reference":[{"key":"ref_1","first-page":"295","article-title":"Differentiation of Duchenne and Becker muscular dystrophy phenotypes with amino- and carboxy-terminal antisera specific for dystrophin","volume":"48","author":"Bulman","year":"1991","journal-title":"Am. J. Hum. Genet."},{"key":"ref_2","first-page":"508","article-title":"A truncated dystrophin lacking the C-terminal domains is localized at the muscle membrane","volume":"50","author":"Helliwell","year":"1992","journal-title":"Am. J. Hum. Genet."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1016\/S1474-4422(03)00585-4","article-title":"Dystrophin and mutations: One gene, several proteins, multiple phenotypes","volume":"2","author":"Muntoni","year":"2003","journal-title":"Lancet Neurol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1186\/s13395-015-0062-6","article-title":"Dp412e: A novel human embryonic dystrophin isoform induced by BMP4 in early differentiated cells","volume":"5","author":"Massourides","year":"2015","journal-title":"Skelet Muscle"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1038\/338509a0","article-title":"Alternative splicing of human dystrophin mRNA generates isoforms at the carboxy terminus","volume":"338","author":"Feener","year":"1989","journal-title":"Nature"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"7205","DOI":"10.1038\/ncomms8205","article-title":"Abnormal splicing switch of DMD\u2019s penultimate exon compromises muscle fibre maintenance in myotonic dystrophy","volume":"6","author":"Rau","year":"2015","journal-title":"Nat. Commun."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"993","DOI":"10.1007\/s00439-015-1581-2","article-title":"Neuronal SH-SY5Y cells use the C-dystrophin promoter coupled with exon 78 skipping and display multiple patterns of alternative splicing including two intronic insertion events","volume":"134","author":"Nishida","year":"2015","journal-title":"Hum. Genet."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"640","DOI":"10.1016\/j.bbrc.2018.11.168","article-title":"Identification of the shortest splice variant of Dp71, together with five known variants, in glioblastoma cells","volume":"508","author":"Rani","year":"2019","journal-title":"Biochem. Biophy. Res. Comm."},{"key":"ref_9","first-page":"100703","article-title":"Schwann cell-specific Dp116 is expressed in glioblastoma cells, revealing two novel DMD gene splicing patterns","volume":"20","author":"Rani","year":"2019","journal-title":"Biochem. Biophys. Rep."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"496","DOI":"10.1038\/nrg3482","article-title":"Alternative cleavage and polyadenylation: Extent, regulation and function","volume":"14","author":"Elkon","year":"2013","journal-title":"Nat. Rev. Genet."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1093\/hmg\/2.5.521","article-title":"Apo-dystrophin-3: A 2.2 kb transcript from the DMD locusencoding the dystrophin glycoprotein binding site","volume":"2","author":"Tinsley","year":"1993","journal-title":"Hum. Mol. Genet."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"926","DOI":"10.1016\/S0960-8966(02)00140-2","article-title":"Survival in Duchenne muscular dystrophy: Improvements in life expectancy since 1967 and the impact of home nocturnal ventilation","volume":"12","author":"Eagle","year":"2002","journal-title":"Neuromuscul. Disord."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.nmd.2010.09.006","article-title":"Duchenne muscular dystrophy: Survival by cardio-respiratory interventions","volume":"21","author":"Ishikawa","year":"2011","journal-title":"Neuromuscul. Disord."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1002\/ppul.23215","article-title":"Cardiac phenotype determines survival in Duchenne muscular dystrophy","volume":"51","author":"Birnkrant","year":"2016","journal-title":"Pediatr. Pulmonol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2799","DOI":"10.1161\/CIRCULATIONAHA.104.528281","article-title":"Genetic predictors and remodeling of dilated cardiomyopathy in muscular dystrophy","volume":"112","author":"Jefferies","year":"2005","journal-title":"Circulation"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1016\/j.amjcard.2014.04.038","article-title":"Left Ventricular Dysfunction in Duchenne Muscular Dystrophy and Genotype","volume":"114","author":"Ashwath","year":"2014","journal-title":"Am. J. Cardiol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Matsuo, M., Awano, H., Matsumoto, M., Nagai, M., Kawaguchi, T., Zhang, Z., and Nishio, H. (2017). Dystrophin Dp116: A yet to Be Investigated Product of the Duchenne Muscular Dystrophy Gene. Genes, 8.","DOI":"10.3390\/genes8100251"},{"key":"ref_18","first-page":"e001782","article-title":"Cardiac dysfunction in Duchenne muscular dystrophy is less frequent in patients with mutations in the dystrophin Dp116 coding region than in other regions","volume":"11","author":"Yamamoto","year":"2018","journal-title":"Circ. Genome Precis. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1016\/0002-8703(52)90209-3","article-title":"Cardiac involvement in progressive muscular dystrophy","volume":"43","author":"Weisenfeld","year":"1952","journal-title":"Am. Heart J."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1016\/0002-8703(78)90358-7","article-title":"Sequence of cardiac changes in Duchenne muscular dystrophy","volume":"95","author":"Heymsfield","year":"1978","journal-title":"Am. Heart J."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1016\/j.pediatrneurol.2008.08.006","article-title":"High incidence of electrocardiogram abnormalities in young patients with duchenne muscular dystrophy","volume":"39","author":"Takami","year":"2008","journal-title":"Pediatr. Neurol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1124","DOI":"10.1161\/01.CIR.35.6.1124","article-title":"The Frank vectorcardiogram and the electrocardiogram in Duchenne progressive muscular dystrophy","volume":"35","author":"Fitch","year":"1967","journal-title":"Circulation"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"853","DOI":"10.1161\/01.CIR.66.4.853","article-title":"Cardiac conduction abnormalities in children with Duchenne\u2019s progressive muscular dystrophy: Electrocardiographic features and morphologic correlates","volume":"66","author":"Sanyal","year":"1982","journal-title":"Circulation"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"462","DOI":"10.1016\/j.nmd.2011.04.005","article-title":"Electrocardiographic abnormalities in very young Duchenne muscular dystrophy patients precede the onset of cardiac dysfunction","volume":"21","author":"James","year":"2011","journal-title":"Neuromuscul. Disord."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2127","DOI":"10.1172\/JCI115244","article-title":"Exon skipping during splicing of dystrophin mRNA precursor due to an intraexon deletion in the dystrophin gene of Duchenne muscular dystrophy kobe","volume":"87","author":"Matsuo","year":"1991","journal-title":"J. Clin. Investig."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1007\/s10038-005-0272-6","article-title":"A novel cryptic exon identified in the 3\u2032 region of intron 2 of the human dystrophin gene","volume":"50","author":"Tran","year":"2005","journal-title":"J. Hum. Genet."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/S1474-4422(09)70271-6","article-title":"Diagnosis and management of Duchenne muscular dystrophy, part 1: Diagnosis, and pharmacological and psychosocial management","volume":"9","author":"Bushby","year":"2010","journal-title":"Lancet Neurol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.echo.2014.10.003","article-title":"Recommendations for cardiac chamber quantification by echocardiography in adults: An update from the American Society of Echocardiography and the European Association of Cardiovascular Imaging","volume":"28","author":"Lang","year":"2015","journal-title":"J. Am. Soc. Echocardiogr."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1214","DOI":"10.1161\/CIRCHEARTFAILURE.113.000125","article-title":"Predictors of disease progression in pediatric dilated cardiomyopathy","volume":"6","author":"Molina","year":"2013","journal-title":"Circ. Heart Fail."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Zhao, J., Kodippili, K., Yue, Y., Hakim, C.H., Wasala, L., Pan, X., Zhang, K., Yang, N.N., Duan, D., and Lai, Y. (2016). Dystrophin contains multiple independent membrane-binding domains. Hum. Mol. Genet.","DOI":"10.1093\/hmg\/ddw210"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/0888-7543(88)90113-9","article-title":"An explanation for the phenotypic differences between patients bearing partial deletions of the DMD locus","volume":"2","author":"Monaco","year":"1988","journal-title":"Genomics"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1080\/15476286.2015.1125074","article-title":"Non-sequential and multi-step splicing of the dystrophin transcript","volume":"13","author":"Gazzoli","year":"2016","journal-title":"RNA Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"12575","DOI":"10.1038\/s41598-017-12981-5","article-title":"Timing and localization of human dystrophin isoform expression provide insights into the cognitive phenotype of Duchenne muscular dystrophy","volume":"7","author":"Doorenweerd","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2766","DOI":"10.1016\/j.celrep.2019.02.049","article-title":"Regulation of Intronic Polyadenylation by PCF11 Impacts mRNA Expression of Long Genes","volume":"26","author":"Wang","year":"2019","journal-title":"Cell Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"346","DOI":"10.3389\/fphys.2012.00346","article-title":"Induced pluripotent stem cell derived cardiomyocytes as models for cardiac arrhythmias","volume":"3","author":"Hoekstra","year":"2012","journal-title":"Front. Physiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1038\/ng0593-82","article-title":"Dystrophin expression in the human retina is required for normal function as defined by electroretinography","volume":"4","author":"Pillers","year":"1993","journal-title":"Nat. Genet"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"562","DOI":"10.1038\/ejhg.2015.135","article-title":"Ocular and neurodevelopmental features of Duchenne muscular dystrophy: A signature of dystrophin function in the central nervous system","volume":"24","author":"Ricotti","year":"2016","journal-title":"Eur. J. Hum. Genet."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"552","DOI":"10.1038\/sj.ejhg.5200488","article-title":"Severe cognitive impairment in DMD: Obvious clinical indication for Dp71 isoform point mutation screening","volume":"8","author":"Moizard","year":"2000","journal-title":"Eur. J. Hum. Genet."},{"key":"ref_39","first-page":"294","article-title":"Expression of Dp71 in Muller glial cells: A comparison with utrophin- and dystrophin-associated proteins","volume":"41","author":"Claudepierre","year":"2000","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1023","DOI":"10.1016\/j.nmd.2017.06.007","article-title":"Patients with Duchenne muscular dystrophy are significantly shorter than those with Becker muscular dystrophy, with the higher incidence of short stature in Dp71 mutated subgroup","volume":"27","author":"Matsumoto","year":"2017","journal-title":"Neuromuscul. Disord."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2125","DOI":"10.1111\/jcmm.14124","article-title":"Electrophysiological abnormalities in induced pluripotent stem cell-derived cardiomyocytes generated from Duchenne muscular dystrophy patients","volume":"23","author":"Eisen","year":"2019","journal-title":"J. Cell. Mol. Med."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1808","DOI":"10.1016\/j.hrthm.2014.05.037","article-title":"From beat rate variability in induced pluripotent stem cell-derived pacemaker cells to heart rate variability in human subjects","volume":"11","author":"Schick","year":"2014","journal-title":"Heart Rhythm"},{"key":"ref_43","first-page":"37","article-title":"Alternative polyadenylation can regulate post-translational membrane localization","volume":"10","author":"Mitra","year":"2015","journal-title":"Trends Cell. Mol. Biol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/10\/3555\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:29:55Z","timestamp":1760174995000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/10\/3555"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,5,18]]},"references-count":43,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2020,5]]}},"alternative-id":["ijms21103555"],"URL":"https:\/\/doi.org\/10.3390\/ijms21103555","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,5,18]]}}}