TAILIEUCHUNG - Báo cáo khoa học: RNA reprogramming of a-mannosidase mRNA sequences in vitro by myxomycete group IC1 and IE ribozymes

Trans-splicing group I ribozymes have been introduced in order to mediate RNA reprogramming (including RNA repair) of therapeutically relevant RNA transcripts. Efficient RNA reprogramming depends on the appropri-ate efficiency of the reaction, and several attempts, including optimization of target recognition and ribozyme catalysis, have been performed. | ềFEBS Journal RNA reprogramming of a-mannosidase mRNA sequences in vitro by myxomycete group IC1 and IE ribozymes Tonje Fiskaa1 Eirik W. Lundblad1 2 J0rn R. Henriksen1 3 Steinar D. Johansen1 and Christer Einvik1 3 1 Department of Molecular Biotechnology RNA Research group Institute of MedicalBiology University of Tromso Norway 2 Department of Microbiology University Hospitalof North Norway Tromso Norway 3 Department of Pediatrics University Hospitalof North Norway Tromso Norway Keywords a-mannosidase mRNA group I intron RNA repair RNA reprogramming trans-splicing Correspondence S. D. Johansen Department of Molecular Biotechnology RNA Research Group Institute of MedicalBiology University of Tromso N-9037 Tromso Norway Fax 47 776 45350 Tel 47 776 45367 E-mail These authors contributed equally to this study Received 17 March 2006 accepted 27 April 2006 doi Trans-splicing group I ribozymes have been introduced in order to mediate RNA reprogramming including RNA repair of therapeutically relevant RNA transcripts. Efficient RNA reprogramming depends on the appropriate efficiency of the reaction and several attempts including optimization of target recognition and ribozyme catalysis have been performed. In most studies the Tetrahymena group IC1 ribozyme has been applied. Here we investigate the potential of group IC1 and group IE intron ribozymes derived from the myxomycetes Didymium and Fuligo in addition to the Tetrahymena ribozyme for RNA reprogramming of a mutated a-mannosi-dase mRNA sequence. Randomized internal guide sequences were introduced for all four ribozymes and used to select accessible sites within isolated mutant a-mannosidase mRNA from mammalian COS-7 cells. Two accessible sites common to all the group I ribozymes were identified and further investigated in RNA reprogramming by trans-splicing analyses. All the myxomycete ribozymes performed the trans-splicing reaction with high fidelity .

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