Use of engineered ribozymes to catalyze chimeric gene assembly.
We report the use of engineered ribozymes to shuffle exon cassettes in vitro. Specifically, we have designed derivatives of a group II intron that insert into selected sites in the human tissue plasminogen activator (t-PA) mRNA. The insertion reaction links t-PA sequences to the group II intron sequences so that trans-splicing reactions catalyzed by the intron can be employed to shuffle the t-PA sequences. We expect these results to be generalizable, so that similar ribozymes can be designed to target any desired 13 nucleotide sequence. In principle, the reactions we describe here should be able to link any RNA molecule to any other RNA molecule at any selected point.
Mikheeva
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Department of Pharmacology and Experimental Therapeutics, Boston University Medical Center, MA 02118, USA.
Jarrell
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KA
D001483:Base Sequence; D017931:DNA Primers; D004815:Epidermal Growth Factor / Q000096:biosynthesis; D005091:Exons; D005353:Fibronectins / Q000096:biosynthesis; D005818:Genetic Engineering / Q000379:methods; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012326:RNA Splicing; D016337:RNA, Catalytic / Q000096:biosynthesis / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D015183:Restriction Mapping; D010959:Tissue Plasminogen Activator / Q000096:biosynthesis*; D014158:Transcription, Genetic
D017931:DNA Primers; D005353:Fibronectins; D016337:RNA, Catalytic; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D004815:Epidermal Growth Factor; D010959:Tissue Plasminogen Activator