The design and optimization of RNA trans-splicing molecules for skin cancer therapy

Christina Gruber, Ulrich Koller, Eva M. Murauer, Stefan Hainzl, Clemens Hüttner, Thomas Kocher, Andrew P. South, Helmut Hintner, Johann W. Bauer

    Research output: Contribution to journalArticlepeer-review

    18 Citations (Scopus)

    Abstract

    Targeting tumor marker genes by RNA trans-splicing is a promising means to induce tumor cell-specific death. Using a screening system we designed RNA trans-splicing molecules (RTM) specifically binding the pre-mRNA of SLCO1B3, a marker gene in epidermolysis bullosa associated squamous cell carcinoma (EB-SCC). Specific trans-splicing, results in the fusion of the endogenous target mRNA of SLCO1B3 and the coding sequence of the suicide gene, provided by the RTM. SLCO1B3-specific RTMs containing HSV-tk were analyzed regarding their trans-splicing potential in a heterologous context using a SLCO1B3 expressing minigene ( SLCO1B3-MG). Expression of the chimeric SLCO1B3-tk was detected by semi-quantitative RT-PCR and Western blot analysis. Cell viability and apoptosis assays confirmed that the RTMs induced suicide gene-mediated apoptosis in SLCO1B3-MG expressing cells. The lead RTM also showed its potential to facilitate a trans-splicing reaction into the endogenous SLCO1B3 pre-mRNA in EB-SCC cells resulting in tk-mediated apoptosis. We assume that the pre-selection of RTMs by our inducible cell-death system accelerates the design of optimal RTMs capable to induce tumor specific cell death in skin cancer cells.
    Original languageEnglish
    Pages (from-to)1056-1068
    Number of pages13
    JournalMolecular Oncology
    Volume7
    Issue number6
    DOIs
    Publication statusPublished - Dec 2013

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