title: Ribozyme-mediated repair of defective mRNA by targeted, trans-splicing.
abstract: Ribozymes can be targeted to cleave specific RNAs, which has led to much interest in their potential as gene inhibitors. Such trans-cleaving ribozymes join a growing list of agents that stop the flow of genetic information. Here we describe a different application of ribozymes for which they may be uniquely suited. By targeted trans-splicing, a ribozyme can replace a defective portion of RNA with a functional sequence. The self-splicing intron from Tetrahymena thermophila was previously shown to mediate trans-splicing of oligonucleotides in vitro. As a model system for messenger RNA repair, this group I intron was re-engineered to regenerate the proper coding capacity of short, truncated lacZ transcripts. Trans-splicing was efficient in vitro and proceeded in Escherichia coli to generate translatable lacZ messages. Targeted trans-splicing represents a general means of altering the sequence of specified transcripts and may provide a new approach to the treatment of many genetic diseases.
authors.affiliation: Howard Hughes Medical Institute, Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309-0215.
authors.forename: B A
authors.initials: BA
authors.lastname: Sullenger
authors.forename: T R
authors.initials: TR
authors.lastname: Cech
mesh_terms: D000818:Animals; D001483:Base Sequence; D016054:DNA, Protozoan; D004926:Escherichia coli / Q000235:genetics; D007438:Introns; D007763:Lac Operon; D008969:Molecular Sequence Data; D012326:RNA Splicing; D016337:RNA, Catalytic / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D016053:RNA, Protozoan; D016808:Tetrahymena thermophila / Q000235:genetics / Q000378:metabolism
chemical_list: D016054:DNA, Protozoan; D016337:RNA, Catalytic; D012333:RNA, Messenger; D016053:RNA, Protozoan