{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T15:40:28Z","timestamp":1787586028946,"version":"build-2736575974"},"reference-count":38,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2018,3,7]],"date-time":"2018-03-07T00:00:00Z","timestamp":1520380800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>In recent years, RNA trans-splicing has emerged as a suitable RNA editing tool for the specific replacement of mutated gene regions at the pre-mRNA level. Although the technology has been successfully applied for the restoration of protein function in various genetic diseases, a higher trans-splicing efficiency is still desired to facilitate its clinical application. Here, we describe a modified, easily applicable, fluorescence-based screening system for the generation and analysis of antisense molecules specifically capable of improving the RNA reprogramming efficiency of a selected KRT14-specific RNA trans-splicing molecule. Using this screening procedure, we identified several antisense RNAs and short rationally designed oligonucleotides, which are able to increase the trans-splicing efficiency. Thus, we assume that besides the RNA trans-splicing molecule, short antisense molecules can act as splicing modulators, thereby increasing the trans-splicing efficiency to a level that may be sufficient to overcome the effects of certain genetic predispositions, particularly those associated with dominantly inherited diseases.<\/jats:p>","DOI":"10.3390\/ijms19030762","type":"journal-article","created":{"date-parts":[[2018,3,7]],"date-time":"2018-03-07T12:55:23Z","timestamp":1520427323000},"page":"762","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["RNA Trans-Splicing Modulation via Antisense Molecule Interference"],"prefix":"10.3390","volume":"19","author":[{"given":"Bernadette","family":"Liemberger","sequence":"first","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Josefina","family":"Pi\u00f1\u00f3n Hofbauer","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Verena","family":"Wally","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia","family":"Arzt","sequence":"additional","affiliation":[{"name":"Laboratory for Immunological and Molecular Cancer Research, Department of Internal Medicine III with Hematology, Medical Oncology, Hemostaseology, Infectious Diseases, Rheumatology, Oncologic Center, Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan","family":"Hainzl","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Kocher","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Murauer","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johann","family":"Bauer","sequence":"additional","affiliation":[{"name":"Department of Dermatology, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4358-7011","authenticated-orcid":false,"given":"Julia","family":"Reichelt","sequence":"additional","affiliation":[{"name":"EB House Austria, Research Program for Molecular Therapy of Genodermatoses, Department of Dermatology, 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, University Hospital of the Paracelsus Medical University, 5020 Salzburg, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2018,3,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1111\/exd.13141","article-title":"RNA-based therapies for genodermatoses","volume":"26","author":"Bornert","year":"2017","journal-title":"Exp. Dermatol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1089\/hum.2013.164","article-title":"Triple trans-splicing adeno-associated virus vectors capable of transferring the coding sequence for full-length dystrophin protein into dystrophic mice","volume":"25","author":"Koo","year":"2014","journal-title":"Hum. Gene Ther."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1038\/nrd3625","article-title":"RNA therapeutics: Beyond RNA interference and antisense oligonucleotides","volume":"11","author":"Kole","year":"2012","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1523\/JNEUROSCI.4489-09.2010","article-title":"Trans-splicing-mediated improvement in a severe mouse model of spinal muscular atrophy","volume":"30","author":"Coady","year":"2010","journal-title":"J. Neurosci."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Lorain, S., Peccate, C., Le Hir, M., and Garcia, L. (2010). Exon Exchange Approach to Repair Duchenne Dystrophin Transcripts. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0010894"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"4715","DOI":"10.1093\/hmg\/ddq405","article-title":"K14 mRNA reprogramming for dominant epidermolysis bullosa simplex","volume":"19","author":"Wally","year":"2010","journal-title":"Hum. Mol. Genet."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"10259","DOI":"10.1093\/nar\/gkx669","article-title":"An RNA-targeted therapy for dystrophic epidermolysis bullosa","volume":"45","author":"Peking","year":"2017","journal-title":"Nucleic Acids Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"568","DOI":"10.1038\/sj.jid.5701152","article-title":"5\u2032 trans-splicing repair of the PLEC1 gene","volume":"128","author":"Wally","year":"2008","journal-title":"J. Investig. Dermatol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"E108","DOI":"10.1093\/nar\/gkr465","article-title":"A novel screening system improves genetic correction by internal exon replacement","volume":"39","author":"Koller","year":"2011","journal-title":"Nucleic Acids Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1176","DOI":"10.1038\/mt.2014.35","article-title":"Cis-splicing and Translation of the Pre-Trans-splicing Molecule Combine With Efficiency in Spliceosome-mediated RNA Trans-splicing","volume":"22","author":"Monjaret","year":"2014","journal-title":"Mol. Ther."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2603","DOI":"10.1093\/hmg\/ddt108","article-title":"Trans-splicing correction of tau isoform imbalance in a mouse model of tau mis-splicing","volume":"22","author":"Avale","year":"2013","journal-title":"Hum. Mol. Genet."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Rindt, H., Yen, P.-F., Thebeau, C.N., Peterson, T.S., Weisman, G.A., and Lorson, C.L. (2012). Replacement of huntingtin exon 1 by trans-splicing. Cell. Mol. Life Sci., 69.","DOI":"10.1007\/s00018-012-1083-5"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/j.tig.2004.12.007","article-title":"The natural history of group I introns","volume":"21","author":"Haugen","year":"2005","journal-title":"Trends Genet."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1261\/rna.028472.111","article-title":"An in vivo selection method to optimize trans-splicing ribozymes","volume":"18","author":"Olson","year":"2012","journal-title":"RNA"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1593","DOI":"10.1126\/science.280.5369.1593","article-title":"Ribozyme-mediated repair of sickle \u03b2-globin mRNAs in erythrocyte precursors","volume":"280","author":"Lan","year":"1998","journal-title":"Science"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1016\/j.ymthe.2005.06.096","article-title":"Specific regression of human cancer cells by ribozyme-mediated targeted replacement of tumor-specific transcript","volume":"12","author":"Kwon","year":"2005","journal-title":"Mol. Ther. J. Am. Soc. Gene Ther."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"357","DOI":"10.7150\/thno.13621","article-title":"Image-aided Suicide Gene Therapy Utilizing Multifunctional hTERT-targeting Adenovirus for Clinical Translation in Hepatocellular Carcinoma","volume":"6","author":"Kim","year":"2016","journal-title":"Theranostics"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"E157","DOI":"10.1038\/mtna.2014.10","article-title":"Considerations for a Successful RNA Trans-splicing Repair of Genetic Disorders","volume":"3","author":"Koller","year":"2014","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"H\u00fcttner, C., Murauer, E.M., Hainzl, S., Kocher, T., Neumayer, A., Reichelt, J., Bauer, J.W., and Koller, U. (2016). Designing Efficient Double RNA trans-Splicing Molecules for Targeted RNA Repair. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17101609"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1103","DOI":"10.1016\/j.jaad.2014.01.903","article-title":"Inherited epidermolysis bullosa: Updated recommendations on diagnosis and classification","volume":"70","author":"Fine","year":"2014","journal-title":"J. Am. Acad. Dermatol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1007\/978-1-62703-227-8_30","article-title":"RNA trans-splicing for genodermatoses","volume":"961","author":"Bauer","year":"2013","journal-title":"Methods Mol. Biol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1089\/hgtb.2012.180","article-title":"A reporter-based screen to identify potent 3\u2019 trans-splicing molecules for endogenous RNA repair","volume":"24","author":"Murauer","year":"2013","journal-title":"Hum. Gene Ther. Methods"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"E287","DOI":"10.1038\/mtna.2016.3","article-title":"A Gene Gun-mediated Nonviral RNA trans-splicing Strategy for COL7A1 Repair","volume":"5","author":"Peking","year":"2016","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"775","DOI":"10.1038\/gt.2016.57","article-title":"Construction and validation of an RNA trans-splicing molecule suitable to repair a large number of COL7A1 mutations","volume":"23","author":"Tockner","year":"2016","journal-title":"Gene Ther."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1038\/mt.2008.234","article-title":"Injection of recombinant human type VII collagen corrects the disease phenotype in a murine model of dystrophic epidermolysis bullosa","volume":"17","author":"Remington","year":"2009","journal-title":"Mol. Ther. J. Am. Soc. Gene Ther."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1016\/0092-8674(92)90314-3","article-title":"The genetic basis of epidermolytic hyperkeratosis: A disorder of differentiation-specific epidermal keratin genes","volume":"70","author":"Cheng","year":"1992","journal-title":"Cell"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"821","DOI":"10.1016\/0092-8674(92)90315-4","article-title":"A leucine\u2014Proline mutation in the H1 subdomain of keratin 1 causes epidermolytic hyperkeratosis","volume":"70","author":"Chipev","year":"1992","journal-title":"Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1179","DOI":"10.3390\/ijms16011179","article-title":"Trans-Splicing Improvement by the Combined Application of Antisense Strategies","volume":"16","author":"Koller","year":"2015","journal-title":"Int. J. Mol. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1007\/978-1-61779-767-5_15","article-title":"Antisense-mediated exon skipping to reframe transcripts","volume":"867","author":"Turczynski","year":"2012","journal-title":"Methods Mol. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2614","DOI":"10.1038\/sj.jid.5700435","article-title":"Targeted skipping of a single exon harboring a premature termination codon mutation: Implications and potential for gene correction therapy for selective dystrophic epidermolysis bullosa patients","volume":"126","author":"Goto","year":"2006","journal-title":"J. Investig. Dermatol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1660","DOI":"10.1073\/pnas.1614775114","article-title":"Efficient in vivo gene editing using ribonucleoproteins in skin stem cells of recessive dystrophic epidermolysis bullosa mouse model","volume":"114","author":"Wu","year":"2017","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Coady, T.H., Baughan, T.D., Shababi, M., Passini, M.A., and Lorson, C.L. (2008). Development of a single vector system that enhances trans-splicing of SMN2 transcripts. PLoS ONE, 3.","DOI":"10.1371\/journal.pone.0003468"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1186\/1750-1172-5-12","article-title":"Inherited epidermolysis bullosa","volume":"5","author":"Fine","year":"2010","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2585","DOI":"10.1016\/j.ymthe.2017.08.015","article-title":"Cut and Paste: Efficient Homology-Directed Repair of a Dominant Negative KRT14 Mutation via CRISPR\/Cas9 Nickases","volume":"25","author":"Kocher","year":"2017","journal-title":"Mol. Ther. J. Am. Soc. Gene Ther."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1158\/1535-7163.MCT-10-0669","article-title":"Spliceosome-mediated RNA trans-splicing facilitates targeted delivery of suicide genes to cancer cells","volume":"10","author":"Gruber","year":"2011","journal-title":"Mol. Cancer Ther."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1056","DOI":"10.1016\/j.molonc.2013.08.005","article-title":"The design and optimization of RNA trans-splicing molecules for skin cancer therapy","volume":"7","author":"Gruber","year":"2013","journal-title":"Mol. Oncol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1083\/jcb.152.3.651","article-title":"An Inducible Mouse Model for Epidermolysis Bullosa Simplex","volume":"152","author":"Cao","year":"2001","journal-title":"J. Cell Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"E67","DOI":"10.1093\/nar\/gkp215","article-title":"Human Splicing Finder: An online bioinformatics tool to predict splicing signals","volume":"37","author":"Desmet","year":"2009","journal-title":"Nucleic Acids Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/19\/3\/762\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T14:57:53Z","timestamp":1760194673000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/19\/3\/762"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,3,7]]},"references-count":38,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2018,3]]}},"alternative-id":["ijms19030762"],"URL":"https:\/\/doi.org\/10.3390\/ijms19030762","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,3,7]]}}}