tailieunhanh - Báo cáo khoa học: Trans-splicing of a mutated glycosylasparaginase mRNA sequence by a group I ribozyme deficient in hydrolysis

RNAreprogramming represents anewconcept incorrecting genetic defects at theRNA , for the technique to be useful for therapy, the level of reprogrammingmust be appropriate. To improve the efficiency of group I ribozyme-mediated RNA reprogramming, when using theTetrahy-menaribozyme, regions complementary to the target RNA have previously been extended in length and accessible sites in the targetRNAs have been identified. | Eur. J. Biochem. 271 4932-4938 2004 FEBS 2004 doi Trafls-splicing of a mutated glycosylasparaginase mRNA sequence by a group I ribozyme deficient in hydrolysis Eirik W. Lundblad1 Peik Haugen1 and Steinar D. Johansen1 2 1 Department of Molecular Biotechnology RNA Research group Institute of Medical Biology University of Troms0 Norway department of Fisheries and Natural Sciences Bode Regional University Norway RNA reprogramming represents a new concept in correcting genetic defects at the RNA level. However for the technique to be useful for therapy the level of reprogramming must be appropriate. To improve the efficiency of group I ribozyme-mediated RNA reprogramming when using the Tetrahy-mena ribozyme regions complementary to the target RNA have previously been extended in length and accessible sites in the target RNAs have been identified. As an alternative to the Tetrahymena model ribozyme the DiGIR2 group I ribozyme derived from a mobile group I intron in rDNA of the myxomycete Didymium iridis represents a new and attractive tool in RNA reprogramming. We reported recently that the deletion of a structural element within the P9 domain of DiGIR2 turns off hydrolysis at the 3 splice site side reaction without affecting self-splicing Haugen P. Andreassen M. Birgisdottir . Johansen . 2004 Eur. J. Biochem. 271 1015-1024 . Here we analyze the potential of the modified ribozyme deficient in hydrolysis at the 3 splice site for application in group I ribozyme-mediated trans-splicing of RNA. The improved ribozyme catalyses both cis-splicing and trans-splicing in vitro of a human glycosylasparaginase mRNA sequence with the same efficiency as the original DiGIR2 ribozyme but without detectable levels of the unwanted hydrolysis. Keywords glycosylasparaginase mRNA group I intron ribozyme hydrolysis RNA reprogramming trans-splicing. Group I ribozyme-mediated RNA reprogramming by trans-splicing has been successfully carried out using the .