Trans-Splicing Improvement by the Combined Application of Antisense Strategies
AbstractSpliceosome-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. View Full-Text
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Koller, U.; Hainzl, S.; Kocher, T.; Hüttner, C.; Klausegger, A.; Gruber, C.; Mayr, E.; Wally, V.; Bauer, J.W.; Murauer, E.M. Trans-Splicing Improvement by the Combined Application of Antisense Strategies. Int. J. Mol. Sci. 2015, 16, 1179-1191.
Koller U, Hainzl S, Kocher T, Hüttner C, Klausegger A, Gruber C, Mayr E, Wally V, Bauer JW, Murauer EM. Trans-Splicing Improvement by the Combined Application of Antisense Strategies. International Journal of Molecular Sciences. 2015; 16(1):1179-1191.Chicago/Turabian Style
Koller, Ulrich; Hainzl, Stefan; Kocher, Thomas; Hüttner, Clemens; Klausegger, Alfred; Gruber, Christina; Mayr, Elisabeth; Wally, Verena; Bauer, Johann W.; Murauer, Eva M. 2015. "Trans-Splicing Improvement by the Combined Application of Antisense Strategies." Int. J. Mol. Sci. 16, no. 1: 1179-1191.