Abstract
RIBOZYMES can be targeted to cleave specific RNAs1–8, which has led to much interest in their potential as gene inhibitors3,9,10. Such fra/is-cleaving ribozymes join a growing list of agents that stop the flow of genetic information11,12. 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 por-tion of RNA with a functional sequence. The self-splicing intron from Tetrahymena thermophila13 was previously shown to mediatetrans-splicing of oligonucleotides in vitro14,15. 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 frans-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.
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Sullenger, B., Cech, T. Ribozyme-mediated repair of defective mRNA by targeted trans-splicing. Nature 371, 619–622 (1994). https://doi.org/10.1038/371619a0
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DOI: https://doi.org/10.1038/371619a0
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