title: Design and characterization of a twin ribozyme for potential repair of a deletion mutation within the oncogenic CTNNB1-ΔS45 mRNA.
abstract: RNA repair is an emerging strategy for gene therapy. Conventional gene therapy typically relies on the addition of the corrected DNA sequence of a defective gene to restore gene function. As an additional option, RNA repair allows alteration of the sequence of endogenous messenger RNAs (mRNAs). mRNA sequence alteration is either facilitated by intracellular spliceosome machinery or by the intrinsic catalytic activity of trans-acting ribozymes. Previously we developed twin ribozymes, derived from the hairpin ribozyme, by tandem duplication and demonstrated their potential for patchwise RNA repair. Herein we describe the development of such a twin ribozyme for potential repair of a deletion mutation in the oncogenic CTNNB1-ΔS45 mRNA. We demonstrate that hairpin ribozyme units within the twin ribozyme can be adapted to efficiently cleave/ligate non-consensus substrates by introduction of compensatory mutations in the ribozyme. Thus, we show the twin ribozyme mediated repair of truncated CTNNB1 transcripts (up to 1000 nt length). Repair of the entire CTNNB1-ΔS45 mRNA, although apparently possible in general, is hampered in vitro by the secondary structure of the transcript.
authors.affiliation: Ernst-Moritz-Arndt Universität Greifswald, Institut für Biochemie, Felix-Hausdorff-Strasse 4, 17478 Greifswald (Germany).
authors.forename: Darko
authors.initials: D
authors.lastname: Balke
authors.forename: Irene
authors.initials: I
authors.lastname: Zieten
authors.forename: Anne
authors.initials: A
authors.lastname: Strahl
authors.forename: Oliver
authors.initials: O
authors.lastname: Müller
authors.forename: Sabine
authors.initials: S
authors.lastname: Müller
mesh_terms: D001483:Base Sequence; D004260:DNA Repair / Q000187:drug effects*; D015195:Drug Design; D017353:Gene Deletion; D007700:Kinetics; D008969:Molecular Sequence Data; D009154:Mutation / Q000187:drug effects*; D059367:RNA Cleavage / Q000187:drug effects; D016337:RNA, Catalytic / Q000138:chemical synthesis* / Q000494:pharmacology*; D012333:RNA, Messenger / Q000187:drug effects*; D051176:beta Catenin / Q000187:drug effects* / Q000235:genetics
chemical_list: C495270:CTNNB1 protein, human; D016337:RNA, Catalytic; D012333:RNA, Messenger; D051176:beta Catenin; C418481:hairpin ribozyme
keywords: RNA repair; gene technology; ribozymes; shape chemistry; β-catenin