{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T01:58:09Z","timestamp":1789351089124,"version":"build-2803163510"},"reference-count":25,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2015,1,6]],"date-time":"2015-01-06T00:00:00Z","timestamp":1420502400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Austrian Science Fund (FWF)","award":["P25304-B19"],"award-info":[{"award-number":["P25304-B19"]}]},{"name":"DEBRA Austria"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Spliceosome-mediated RNA trans-splicing has become an emergent tool for  the repair of mutated pre-mRNAs in the treatment of genetic diseases. RNA trans-splicing molecules (RTMs) are designed to induce a specific trans-splicing reaction via a binding domain for a respective target pre-mRNA region. A previously established reporter-based screening system allows us to analyze the impact of various factors on the RTM  trans-splicing efficiency in vitro. Using this system, we are further able to investigate the potential of antisense RNAs (AS RNAs), presuming to improve the trans-splicing efficiency of a selected RTM, specific for intron 102 of COL7A1. Mutations in the COL7A1 gene underlie the dystrophic subtype of the skin blistering disease epidermolysis bullosa (DEB). We have shown that co-transfections of the RTM and a selected AS RNA, interfering with competitive splicing elements on a COL7A1-minigene (COL7A1-MG), lead to a significant increase of the RNA trans-splicing efficiency. Thereby, accurate trans-splicing between  the RTM and the COL7A1-MG is represented by the restoration of full-length green fluorescent protein GFP on mRNA and protein level. This mechanism can be crucial for the improvement of an RTM-mediated correction, especially in cases where a high trans-splicing efficiency is required.<\/jats:p>","DOI":"10.3390\/ijms16011179","type":"journal-article","created":{"date-parts":[[2015,1,6]],"date-time":"2015-01-06T09:11:06Z","timestamp":1420535466000},"page":"1179-1191","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Trans-Splicing Improvement by the Combined Application  of Antisense Strategies"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6285-1789","authenticated-orcid":false,"given":"Ulrich","family":"Koller","sequence":"first","affiliation":[{"name":"Department of Dermatology and EB House Austria, Paracelsus Medical University, Salzburg 5020, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan","family":"Hainzl","sequence":"additional","affiliation":[{"name":"Department of Dermatology and EB House Austria, Paracelsus Medical University, Salzburg 5020, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Kocher","sequence":"additional","affiliation":[{"name":"Department of Dermatology and EB House Austria, Paracelsus Medical University, Salzburg 5020, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Clemens","family":"H\u00fcttner","sequence":"additional","affiliation":[{"name":"Department of Dermatology and EB House Austria, Paracelsus Medical University, Salzburg 5020, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alfred","family":"Klausegger","sequence":"additional","affiliation":[{"name":"Department of Dermatology and EB House Austria, Paracelsus Medical University, Salzburg 5020, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christina","family":"Gruber","sequence":"additional","affiliation":[{"name":"Department of Dermatology and EB House Austria, Paracelsus Medical University, Salzburg 5020, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisabeth","family":"Mayr","sequence":"additional","affiliation":[{"name":"Department of Dermatology and EB House Austria, Paracelsus Medical University, Salzburg 5020, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Verena","family":"Wally","sequence":"additional","affiliation":[{"name":"Department of Dermatology and EB House Austria, Paracelsus Medical University, Salzburg 5020, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johann","family":"Bauer","sequence":"additional","affiliation":[{"name":"Department of Dermatology and EB House Austria, Paracelsus Medical University, Salzburg 5020, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Murauer","sequence":"additional","affiliation":[{"name":"Department of Dermatology and EB House Austria, Paracelsus Medical University, Salzburg 5020, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2015,1,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1038\/nm900","article-title":"Phenotype correction of hemophilia A mice by spliceosome-mediated RNA trans-splicing","volume":"9","author":"Chao","year":"2003","journal-title":"Nat. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1290","DOI":"10.1261\/rna.5101903","article-title":"5' Exon replacement and repair by spliceosome-mediated RNA trans-splicing","volume":"9","author":"Mansfield","year":"2003","journal-title":"RNA"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"568","DOI":"10.1038\/sj.jid.5701152","article-title":"5' Trans-splicing repair of the PLEC1 gene","volume":"128","author":"Wally","year":"2008","journal-title":"J. Investig. Dermatol."},{"key":"ref_4","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_5","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1038\/jid.2010.249","article-title":"Functional correction of type VII collagen expression in dystrophic epidermolysis bullosa","volume":"131","author":"Murauer","year":"2011","journal-title":"J. Investig. Dermatol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1959","DOI":"10.1038\/jid.2012.101","article-title":"Spliceosome-mediated trans-splicing: The therapeutic cut and paste","volume":"132","author":"Wally","year":"2012","journal-title":"J. Investig. Dermatol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1016\/j.det.2010.02.003","article-title":"Gene therapy for recessive dystrophic epidermolysis bullosa","volume":"28","author":"Titeux","year":"2010","journal-title":"Dermatol. Clin."},{"key":"ref_8","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_9","doi-asserted-by":"crossref","unstructured":"Plaseska-Karanfilska, Dijana (2011). Human Genetic Diseases, InTech.","DOI":"10.5772\/904"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1089\/hgtb.2012.180","article-title":"A reporter-based screen to identify potent 3' trans-splicing molecules for endogenous RNA repair","volume":"24","author":"Murauer","year":"2013","journal-title":"Hum. Gene Ther. Methods"},{"key":"ref_11","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_12","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_13","doi-asserted-by":"crossref","first-page":"e3468","DOI":"10.1371\/journal.pone.0003468","article-title":"Development of a single vector system that enhances trans-splicing of SMN2 transcripts","volume":"3","author":"Coady","year":"2008","journal-title":"PLoS One"},{"key":"ref_14","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_15","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_16","doi-asserted-by":"crossref","unstructured":"Fine, J.D., and Hintner, H. (2008). Life with Epidermolysis Bullosa, Springer Wien.","DOI":"10.1007\/978-3-211-79271-1"},{"key":"ref_17","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_18","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_19","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_20","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1038\/6986","article-title":"Spliceosome-mediated RNA trans-splicing as a tool for gene therapy","volume":"17","author":"Puttaraju","year":"1999","journal-title":"Nat. Biotechnol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1477","DOI":"10.1038\/sj.gt.3302596","article-title":"Gene therapy progress and prospects: Reprograming gene expression by trans-splicing","volume":"12","author":"Mitchell","year":"2005","journal-title":"Gene Ther."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1111\/1523-1747.ep12265460","article-title":"Evaluation of anchoring fibrils and other components of the dermal-epidermal junction in dystrophic epidermolysis bullosa by a quantitative ultrastructural technique","volume":"84","author":"Tidman","year":"1985","journal-title":"J. Investig. Dermatol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1669","DOI":"10.1172\/JCI34292","article-title":"A hypomorphic mouse model of dystrophic epidermolysis bullosa reveals mechanisms of disease and response to fibroblast therapy","volume":"118","author":"Fritsch","year":"2008","journal-title":"J. Clin. Investig."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"30248","DOI":"10.1074\/jbc.M109.045294","article-title":"Dominant negative effects of COL7A1 mutations can be rescued by controlled over-expression of normal collagen VII","volume":"284","author":"Fritsch","year":"2009","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1083\/jcb.152.3.651","article-title":"An inducible mouse model for epidermolysis bullosa simplex: Implications for gene therapy","volume":"152","author":"Cao","year":"2001","journal-title":"J. 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