Tagging ribozyme reaction sites to follow trans-splicing in mammalian cells.

Journal Article (Journal Article)

In mammalian cells, genetic instructions are usually revised by RNA splicing before they are translated to proteins. Here we demonstrate that a trans-splicing group I ribozyme can be employed to intentionally modify the sequence of targeted transcripts in tissue culture cells. By analyzing the ribozyme reaction products, we demonstrate that targeted trans-splicing can proceed in murine fibroblasts with high fidelity, providing direct evidence that ribozymes function as anticipated in a therapeutically relevant setting. Trans-splicing is not very specific however, and the ribozyme reacted with and tagged a variety of cellular transcripts with its 3' exon sequence. RNA tagging provides a unique approach to study RNA catalysis in mammalian cells. Such analysis should facilitate the logical development of safe, therapeutic ribozymes that can repair mutant RNAs associated with a variety of inherited diseases.

Full Text

Duke Authors

Cited Authors

  • Jones, JT; Lee, SW; Sullenger, BA

Published Date

  • June 1996

Published In

Volume / Issue

  • 2 / 6

Start / End Page

  • 643 - 648

PubMed ID

  • 8640554

International Standard Serial Number (ISSN)

  • 1078-8956

Digital Object Identifier (DOI)

  • 10.1038/nm0696-643


  • eng

Conference Location

  • United States