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Among various treatments available for DMD, antisense oligonucleotides (ASOs) mediated exon skipping is a promising therapeutic approach. For successful treatments, however, it is requisite to rigorously optimise oligonucleotide chemistries as well as chemical modifications of ASOs. To achieve this, here, we aim to develop a novel enhanced green fluorescence protein (EGFP)-based reporter assay system that allows us to perform efficient and high-throughput screenings for ASOs. We design a new expression vector with a CAG promoter to detect the EGFP fluorescence only when skipping of <jats:italic>mdx<\/jats:italic>-type exon 23 is induced by ASOs. Then, an accurate screening was successfully conducted in C57BL\/6 primary myotubes using phosphorodiamidate morpholino oligomer or locked nucleic acids (LNA)\/2\u2032-OMe mixmers with different extent of LNA inclusion. We accordingly generated a novel transgenic mouse model with this EGFP expression vector (EGFP-<jats:italic>mdx<\/jats:italic>23 Tg). Finally, we confirmed that the EGFP-<jats:italic>mdx<\/jats:italic>23 Tg provided a highly sensitive platform to check the effectiveness as well as the biodistribution of ASOs for exon skipping therapy. Thus, the assay system provides a simple yet highly sensitive platform to optimise oligonucleotide chemistries as well as chemical modifications of ASOs.<\/jats:p>","DOI":"10.1038\/s41598-020-67077-4","type":"journal-article","created":{"date-parts":[[2020,6,22]],"date-time":"2020-06-22T10:03:36Z","timestamp":1592820216000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Novel EGFP reporter cell and mouse models for sensitive imaging and quantification of exon skipping"],"prefix":"10.1038","volume":"10","author":[{"given":"Yuko","family":"Hara","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoshitaka","family":"Mizobe","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4105-9127","authenticated-orcid":false,"given":"Yukiko U.","family":"Inoue","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yasumasa","family":"Hashimoto","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8172-4339","authenticated-orcid":false,"given":"Norio","family":"Motohashi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoshiaki","family":"Masaki","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kohji","family":"Seio","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4780-6290","authenticated-orcid":false,"given":"Shin\u2019ichi","family":"Takeda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tetsuya","family":"Nagata","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matthew J. A.","family":"Wood","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takayoshi","family":"Inoue","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9038-7506","authenticated-orcid":false,"given":"Yoshitsugu","family":"Aoki","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2020,6,22]]},"reference":[{"key":"67077_CR1","doi-asserted-by":"publisher","first-page":"482","DOI":"10.1016\/j.nmd.2014.03.008","volume":"24","author":"JK Mah","year":"2014","unstructured":"Mah, J. K. et al. A systematic review and meta-analysis on the epidemiology of Duchenne and Becker muscular dystrophy. Neuromuscul Disord 24, 482\u2013491 (2014).","journal-title":"Neuromuscul Disord"},{"key":"67077_CR2","doi-asserted-by":"crossref","unstructured":"Hoffman, E. P. Pharmacotherapy of Duchenne Muscular Dystrophy. Handb Exp Pharmacol [Epub ahead of print] (2019).","DOI":"10.1007\/164_2019_256"},{"key":"67077_CR3","doi-asserted-by":"publisher","first-page":"722","DOI":"10.1097\/WCO.0000000000000739","volume":"32","author":"MA Waldrop","year":"2019","unstructured":"Waldrop, M. A. & Flanigan, K. M. Update in Duchenne and Becker muscular dystrophy. Curr Opin Neurol 32, 722\u2013727 (2019).","journal-title":"Curr Opin Neurol"},{"key":"67077_CR4","doi-asserted-by":"publisher","first-page":"R163","DOI":"10.1093\/hmg\/ddy171","volume":"27","author":"L Echevarria","year":"2018","unstructured":"Echevarria, L., Aupy, P. & Goyenvalle, A. Exon-skipping advances for Duchenne muscular dystrophy. Hum Mol Genet 27, R163\u2013r172 (2018).","journal-title":"Hum Mol Genet"},{"key":"67077_CR5","doi-asserted-by":"publisher","first-page":"251","DOI":"10.1089\/nat.2017.0682","volume":"27","author":"A Aartsma-Rus","year":"2017","unstructured":"Aartsma-Rus, A. et al. Development of Exon Skipping Therapies for Duchenne Muscular Dystrophy: A Critical Review and a Perspective on the Outstanding Issues. Nucleic Acid Ther 27, 251\u2013259 (2017).","journal-title":"Nucleic Acid Ther"},{"key":"67077_CR6","doi-asserted-by":"publisher","DOI":"10.1186\/s13023-017-0631-3","volume":"12","author":"S Ryder","year":"2017","unstructured":"Ryder, S. et al. The burden, epidemiology, costs and treatment for Duchenne muscular dystrophy: an evidence review. Orphanet J Rare Dis 12, 79 (2017).","journal-title":"Orphanet J Rare Dis"},{"key":"67077_CR7","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1089\/nat.2013.0461","volume":"24","author":"K Siva","year":"2014","unstructured":"Siva, K., Covello, G. & Denti, M. A. Exon-skipping antisense oligonucleotides to correct missplicing in neurogenetic diseases. 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