{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T05:38:49Z","timestamp":1792820329021,"version":"4.5.0"},"reference-count":60,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2024,1,7]],"date-time":"2024-01-07T00:00:00Z","timestamp":1704585600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"DEBRA Austria","award":["20204-WISS\/225\/241\/17\u20132021"],"award-info":[{"award-number":["20204-WISS\/225\/241\/17\u20132021"]}]},{"name":"Land Salzburg","award":["20204-WISS\/225\/241\/17\u20132021"],"award-info":[{"award-number":["20204-WISS\/225\/241\/17\u20132021"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Antisense oligonucleotides (ASOs) represent an emerging therapeutic platform for targeting genetic diseases by influencing various aspects of (pre-)mRNA biology, such as splicing, stability, and translation. In this study, we investigated the potential of modulating the splicing pattern in recessive dystrophic epidermolysis bullosa (RDEB) patient cells carrying a frequent genomic variant (c.425A &gt; G) that disrupts splicing in the COL7A1 gene by using short 2\u2032-O-(2-Methoxyethyl) oligoribo-nucleotides (2\u2032-MOE ASOs). COL7A1-encoded type VII collagen (C7) forms the anchoring fibrils within the skin that are essential for the attachment of the epidermis to the underlying dermis. As such, gene variants of COL7A1 leading to functionally impaired or absent C7 manifest in the form of extensive blistering and wounding. The severity of the disease pattern warrants the development of novel therapies for patients. The c.425A &gt; G variant at the COL7A1 exon 3\/intron 3 junction lowers the efficiency of splicing at this junction, resulting in non-functional C7 transcripts. However, we found that correct splicing still occurs, albeit at a very low level, highlighting an opportunity for intervention by modulating the splicing reaction. We therefore screened 2\u2032-MOE ASOs that bind along the COL7A1 target region ranging from exon 3 to the intron 3\/exon 4 junction for their ability to modulate splicing. We identified ASOs capable of increasing the relative levels of correctly spliced COL7A1 transcripts by RT-PCR, sqRT-PCR, and ddPCR. Furthermore, RDEB-derived skin equivalents treated with one of the most promising ASOs exhibited an increase in full-length C7 expression and its accurate deposition along the basement membrane zone (BMZ).<\/jats:p>","DOI":"10.3390\/ijms25020761","type":"journal-article","created":{"date-parts":[[2024,1,8]],"date-time":"2024-01-08T07:59:20Z","timestamp":1704700760000},"page":"761","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Splicing Modulation via Antisense Oligonucleotides in Recessive Dystrophic Epidermolysis Bullosa"],"prefix":"10.3390","volume":"25","author":[{"given":"Stefan","family":"Hainzl","sequence":"first","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology and Allergology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-2372-5848","authenticated-orcid":false,"given":"Lisa","family":"Trattner","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology and Allergology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1056-362X","authenticated-orcid":false,"given":"Bernadette","family":"Liemberger","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology and Allergology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5309-1936","authenticated-orcid":false,"given":"Johannes","family":"Bischof","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology and Allergology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3424-3049","authenticated-orcid":false,"given":"Thomas","family":"Kocher","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology and Allergology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Ablinger","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology and Allergology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4666-2240","authenticated-orcid":false,"given":"Alexander","family":"Nystr\u00f6m","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Medical Faculty, Medical Center\u2014University of Freiburg, 79110 Freiburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Astrid","family":"Obermayer","sequence":"additional","affiliation":[{"name":"Core Facility of Electron Microscopy, Department of Environment & Biodiversity, Paris Lodron University Salzburg (PLUS Salzburg), 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alfred","family":"Klausegger","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology and Allergology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8232-5068","authenticated-orcid":false,"given":"Christina","family":"Guttmann-Gruber","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology and Allergology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8705-3890","authenticated-orcid":false,"given":"Verena","family":"Wally","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology and Allergology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johann W.","family":"Bauer","sequence":"additional","affiliation":[{"name":"Department of Dermatology and Allergology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8558-9031","authenticated-orcid":false,"given":"Josefina Pi\u00f1\u00f3n","family":"Hofbauer","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology and Allergology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6285-1789","authenticated-orcid":false,"given":"Ulrich","family":"Koller","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology and Allergology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,1,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1002\/wrna.1158","article-title":"Targeting RNA splicing for disease therapy","volume":"4","author":"Havens","year":"2013","journal-title":"Wiley Interdiscip. Rev. RNA"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1038\/s12276-022-00757-5","article-title":"RNA therapy: Rich history, various applications and unlimited future prospects","volume":"54","author":"Kim","year":"2022","journal-title":"Exp. Mol. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1002\/wrna.1347","article-title":"mRNA trans-splicing in gene therapy for genetic diseases","volume":"7","author":"Berger","year":"2016","journal-title":"Wiley Interdiscip. Rev. RNA"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"644","DOI":"10.1038\/s41419-022-05075-2","article-title":"RNA-based therapeutics: An overview and prospectus","volume":"13","author":"Zhu","year":"2022","journal-title":"Cell Death Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2573","DOI":"10.1016\/j.ymthe.2017.07.005","article-title":"COL7A1 Editing via CRISPR\/Cas9 in Recessive Dystrophic Epidermolysis Bullosa","volume":"25","author":"Hainzl","year":"2017","journal-title":"Mol. Ther."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3501","DOI":"10.1038\/s41467-020-17093-9","article-title":"Antisense oligonucleotide modulation of non-productive alternative splicing upregulates gene expression","volume":"11","author":"Lim","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Hua, Y., Vickers, T.A., Baker, B.F., Bennett, C.F., and Krainer, A.R. (2007). Enhancement of SMN2 exon 7 inclusion by antisense oligonucleotides targeting the exon. PLoS Biol., 5.","DOI":"10.1371\/journal.pbio.0050073"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"630","DOI":"10.1038\/s41565-021-00898-0","article-title":"The current landscape of nucleic acid therapeutics","volume":"16","author":"Kulkarni","year":"2021","journal-title":"Nat. Nanotechnol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1007\/978-1-61779-767-5_20","article-title":"Antisense-mediated exon inclusion","volume":"867","author":"Hua","year":"2012","journal-title":"Methods Mol. Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1228","DOI":"10.1038\/nbt759","article-title":"Systemically delivered antisense oligomers upregulate gene expression in mouse tissues","volume":"20","author":"Sazani","year":"2002","journal-title":"Nat. Biotechnol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Sogaard, P.P., Lind, M., Christiansen, C.R., Petersson, K., Clauss, A., and Caffarel-Salvador, E. (2021). Future Perspectives of Oral Delivery of Next Generation Therapies for Treatment of Skin Diseases. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13101722"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"614","DOI":"10.1111\/bjd.18921","article-title":"Consensus reclassification of inherited epidermolysis bullosa and other disorders with skin fragility","volume":"183","author":"Has","year":"2020","journal-title":"Br. J. Dermatol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1016\/j.matbio.2018.04.001","article-title":"Epidermolysis bullosa: Molecular pathology of connective tissue components in the cutaneous basement membrane zone","volume":"71","author":"Has","year":"2018","journal-title":"Matrix Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"737272","DOI":"10.3389\/fgene.2021.737272","article-title":"Dystrophic Epidermolysis Bullosa: Secondary Disease Mechanisms and Disease Modifiers","volume":"12","author":"Nystrom","year":"2021","journal-title":"Front. Genet."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"879","DOI":"10.1111\/j.1365-2133.2005.06542.x","article-title":"High frequency of the 425A\u2192G splice-site mutation and novel mutations of the COL7A1 gene in central Europe: Significance for future mutation detection strategies in dystrophic epidermolysis bullosa","volume":"152","author":"Csikos","year":"2005","journal-title":"Br. J. Dermatol."},{"key":"ref_16","first-page":"292","article-title":"Identification of two splicing mutations in the collagen type VII gene (COL7A1) of a patient affected by the localisata variant of recessive dystrophic epidermolysis bullosa","volume":"59","author":"Gardella","year":"1996","journal-title":"Am. J. Hum. Genet."},{"key":"ref_17","unstructured":"FDA (2023, November 18). FDA Approves First Drug for Spinal Muscular Atrophy, Available online: https:\/\/www.fda.gov\/news-events\/press-announcements\/fda-approves-first-drug-spinal-muscular-atrophy."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/0092-8674(95)90460-3","article-title":"Identification and characterization of a spinal muscular atrophy-determining gene","volume":"80","author":"Lefebvre","year":"1995","journal-title":"Cell"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1177","DOI":"10.1093\/hmg\/8.7.1177","article-title":"A single nucleotide difference that alters splicing patterns distinguishes the SMA gene SMN1 from the copy gene SMN2","volume":"8","author":"Monani","year":"1999","journal-title":"Hum. Mol. Genet."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"45476","DOI":"10.1074\/jbc.M107632200","article-title":"Modulation of survival motor neuron pre-mRNA splicing by inhibition of alternative 3\u2032 splice site pairing","volume":"276","author":"Lim","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.omtn.2019.08.014","article-title":"Combining Engineered U1 snRNA and Antisense Oligonucleotides to Improve the Treatment of a BBS1 Splice Site Mutation","volume":"18","author":"Breuel","year":"2019","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"W296","DOI":"10.1093\/nar\/gky427","article-title":"SWISS-MODEL: Homology modelling of protein structures and complexes","volume":"46","author":"Waterhouse","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1167","DOI":"10.1261\/rna.2154310","article-title":"An antisense microwalk reveals critical role of an intronic position linked to a unique long-distance interaction in pre-mRNA splicing","volume":"16","author":"Singh","year":"2010","journal-title":"RNA"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1016\/j.ijpharm.2016.01.042","article-title":"An elastic liposomal formulation for RNAi-based topical treatment of skin disorders: Proof-of-concept in the treatment of psoriasis","volume":"500","author":"Desmet","year":"2016","journal-title":"Int. J. Pharm."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Ablinger, M., Lettner, T., Friedl, N., Potocki, H., Palmetzhofer, T., Koller, U., Illmer, J., Liemberger, B., Hainzl, S., and Klausegger, A. (2021). Personalized Development of Antisense Oligonucleotides for Exon Skipping Restores Type XVII Collagen Expression in Junctional Epidermolysis Bullosa. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22073326"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1038\/nature24487","article-title":"Regeneration of the entire human epidermis using transgenic stem cells","volume":"551","author":"Hirsch","year":"2017","journal-title":"Nature"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.stemcr.2013.11.001","article-title":"Long-term stability and safety of transgenic cultured epidermal stem cells in gene therapy of junctional epidermolysis bullosa","volume":"2","author":"Carulli","year":"2014","journal-title":"Stem Cell Rep."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"780","DOI":"10.1038\/s41591-022-01737-y","article-title":"In vivo topical gene therapy for recessive dystrophic epidermolysis bullosa: A phase 1 and 2 trial","volume":"28","author":"Gurevich","year":"2022","journal-title":"Nat. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2211","DOI":"10.1056\/NEJMoa2206663","article-title":"Trial of Beremagene Geperpavec (B-VEC) for Dystrophic Epidermolysis Bullosa","volume":"387","author":"Guide","year":"2022","journal-title":"N. Engl. J. Med."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1221","DOI":"10.1016\/j.jid.2018.11.036","article-title":"Gene Therapy for Epidermolysis Bullosa","volume":"139","author":"Marinkovich","year":"2019","journal-title":"J. Investig. Dermatol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1175","DOI":"10.1007\/s13555-021-00561-5","article-title":"Clinical Perspectives of Gene-Targeted Therapies for Epidermolysis Bullosa","volume":"11","author":"Welponer","year":"2021","journal-title":"Dermatol. Ther."},{"key":"ref_32","unstructured":"FDA (2023, November 27). VYJUVEK, Available online: https:\/\/www.fda.gov\/vaccines-blood-biologics\/vyjuvek."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"232","DOI":"10.31662\/jmaj.2021-0019","article-title":"Antisense Oligonucleotide-Mediated Exon-skipping Therapies: Precision Medicine Spreading from Duchenne Muscular Dystrophy","volume":"4","author":"Matsuo","year":"2021","journal-title":"JMA J."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1146\/annurev-pharmtox-010818-021050","article-title":"Therapeutic Oligonucleotides: State of the Art","volume":"59","author":"Smith","year":"2019","journal-title":"Annu. Rev. Pharmacol. Toxicol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1016\/j.jid.2020.08.018","article-title":"QR-313, an Antisense Oligonucleotide, Shows Therapeutic Efficacy for Treatment of Dominant and Recessive Dystrophic Epidermolysis Bullosa: A Preclinical Study","volume":"141","author":"Bornert","year":"2021","journal-title":"J. Investig. Dermatol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"e379","DOI":"10.1038\/mtna.2016.87","article-title":"Antisense Oligonucleotide-mediated Exon Skipping as a Systemic Therapeutic Approach for Recessive Dystrophic Epidermolysis Bullosa","volume":"5","author":"Bremer","year":"2016","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2387","DOI":"10.1016\/j.jid.2016.07.029","article-title":"Targeted Exon Skipping Restores Type VII Collagen Expression and Anchoring Fibril Formation in an In Vivo RDEB Model","volume":"136","author":"Turczynski","year":"2016","journal-title":"J. Investig. Dermatol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1038\/nsb887","article-title":"Correction of disease-associated exon skipping by synthetic exon-specific activators","volume":"10","author":"Cartegni","year":"2003","journal-title":"Nat. Struct. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"834","DOI":"10.1016\/j.ajhg.2008.01.014","article-title":"Antisense masking of an hnRNP A1\/A2 intronic splicing silencer corrects SMN2 splicing in transgenic mice","volume":"82","author":"Hua","year":"2008","journal-title":"Am. J. Hum. Genet."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1634","DOI":"10.1101\/gad.1941310","article-title":"Antisense correction of SMN2 splicing in the CNS rescues necrosis in a type III SMA mouse model","volume":"24","author":"Hua","year":"2010","journal-title":"Genes Dev."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"72ra18","DOI":"10.1126\/scitranslmed.3001777","article-title":"Antisense oligonucleotides delivered to the mouse CNS ameliorate symptoms of severe spinal muscular atrophy","volume":"3","author":"Passini","year":"2011","journal-title":"Sci. Transl. Med."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1080\/17425247.2018.1473375","article-title":"Recent preclinical and clinical advances in oligonucleotide conjugates","volume":"15","author":"Craig","year":"2018","journal-title":"Expert Opin. Drug Deliv."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Huang, S., Hao, X.Y., Li, Y.J., Wu, J.Y., Xiang, D.X., and Luo, S. (2022). Nonviral delivery systems for antisense oligonucleotide therapeutics. Biomater. Res., 26.","DOI":"10.1186\/s40824-022-00292-4"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"423","DOI":"10.1007\/s40259-017-0243-4","article-title":"Advances in the Application and Impact of MicroRNAs as Therapies for Skin Disease","volume":"31","author":"Lawrence","year":"2017","journal-title":"BioDrugs"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2075","DOI":"10.1016\/j.ymthe.2017.06.002","article-title":"RNase H1-Dependent Antisense Oligonucleotides Are Robustly Active in Directing RNA Cleavage in Both the Cytoplasm and the Nucleus","volume":"25","author":"Liang","year":"2017","journal-title":"Mol. Ther."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"8277","DOI":"10.1093\/nar\/gkab599","article-title":"Golgi-58K can re-localize to late endosomes upon cellular uptake of PS-ASOs and facilitates endosomal release of ASOs","volume":"49","author":"Liang","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"455","DOI":"10.2147\/TCRM.S386923","article-title":"Innovations in the Treatment of Dystrophic Epidermolysis Bullosa (DEB): Current Landscape and Prospects","volume":"19","author":"Hou","year":"2023","journal-title":"Ther. Clin. Risk Manag."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"e1594","DOI":"10.1002\/wrna.1594","article-title":"The powerful world of antisense oligonucleotides: From bench to bedside","volume":"11","author":"Quemener","year":"2020","journal-title":"Wiley Interdiscip. Rev. RNA"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"eabb6049","DOI":"10.1126\/sciadv.abb6049","article-title":"Treatment of psoriasis with NFKBIZ siRNA using topical ionic liquid formulations","volume":"6","author":"Mandal","year":"2020","journal-title":"Sci. Adv."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1038\/s41573-020-0075-7","article-title":"Advances in oligonucleotide drug delivery","volume":"19","author":"Roberts","year":"2020","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"2853","DOI":"10.1093\/nar\/gkaa126","article-title":"Comparison of the efficacy of MOE and PMO modifications of systemic antisense oligonucleotides in a severe SMA mouse model","volume":"48","author":"Sheng","year":"2020","journal-title":"Nucleic Acids Res."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"10505","DOI":"10.1128\/MCB.24.24.10505-10514.2004","article-title":"Influence of RNA secondary structure on the pre-mRNA splicing process","volume":"24","author":"Buratti","year":"2004","journal-title":"Mol. Cell Biol."},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Ong, A.A.L., Tan, J., Bhadra, M., Dezanet, C., Patil, K.M., Chong, M.S., Kierzek, R., Decout, J.L., Roca, X., and Chen, G. (2019). RNA Secondary Structure-Based Design of Antisense Peptide Nucleic Acids for Modulating Disease-Associated Aberrant Tau Pre-mRNA Alternative Splicing. Molecules, 24.","DOI":"10.3390\/molecules24163020"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1069","DOI":"10.1016\/j.ymthe.2017.03.023","article-title":"FDA-Approved Oligonucleotide Therapies in 2017","volume":"25","author":"Stein","year":"2017","journal-title":"Mol. Ther."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1089\/nat.2016.0657","article-title":"FDA Approves Eteplirsen for Duchenne Muscular Dystrophy: The Next Chapter in the Eteplirsen Saga","volume":"27","author":"Krieg","year":"2017","journal-title":"Nucleic Acid. Ther."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1358\/dot.2019.55.10.3045038","article-title":"Viltolarsen for the treatment of Duchenne muscular dystrophy","volume":"55","author":"Roshmi","year":"2019","journal-title":"Drugs Today"},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Crooke, S.T., Liang, X.H., Crooke, R.M., Baker, B.F., and Geary, R.S. (2021). Antisense drug discovery and development technology considered in a pharmacological context. Biochem. Pharmacol., 189.","DOI":"10.1016\/j.bcp.2020.114196"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1038\/nmeth.2089","article-title":"NIH Image to ImageJ: 25 years of image analysis","volume":"9","author":"Schneider","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_59","doi-asserted-by":"crossref","unstructured":"Bornert, O., Kocher, T., Gretzmeier, C., Liemberger, B., Hainzl, S., Koller, U., and Nystrom, A. (2019). Generation of rabbit polyclonal human and murine collagen VII monospecific antibodies: A useful tool for dystrophic epidermolysis bullosa therapy studies. Matrix Biol. Plus, 4.","DOI":"10.1016\/j.mbplus.2019.100017"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"1985","DOI":"10.1016\/j.jid.2020.02.012","article-title":"Predictable CRISPR\/Cas9-Mediated COL7A1 Reframing for Dystrophic Epidermolysis Bullosa","volume":"140","author":"Kocher","year":"2020","journal-title":"J. Investig. Dermatol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/2\/761\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T13:41:38Z","timestamp":1760103698000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/2\/761"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,1,7]]},"references-count":60,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2024,1]]}},"alternative-id":["ijms25020761"],"URL":"https:\/\/doi.org\/10.3390\/ijms25020761","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,1,7]]}}}