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Nucleic Acids Research 2005 33(4):1240-1248; doi:10.1093/nar/gki278
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Published online 24 February 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
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Article

Target-dependent on/off switch increases ribozyme fidelity

Lucien Junior Bergeron and Jean-Pierre Perreault*

RNA Group/Groupe ARN, Département de Biochimie, Faculté de Médecine, Université de Sherbrooke Sherbrooke, Québec, J1H 5N4, Canada

*To whom correspondence should be addressed. Tel: +1 819 564 5310; Fax: +1 819 564 5340; Email: Jean-Pierre.Perreault{at}USherbrooke.ca

Received January 4, 2005. Revised February 11, 2005. Accepted February 11, 2005.

Ribozymes, RNA molecules that catalyze the cleavage of RNA substrates, provide an interesting alternative to the RNA interference (RNAi) approach to gene inactivation, especially given the fact that RNAi seems to trigger an immunological response. Unfortunately, the limited substrate specificity of ribozymes is considered to be a significant hurdle in their development as molecular tools. Here, we report the molecular engineering of a ribozyme possessing a new biosensor module that switches the cleavage activity from ‘off’ (a ‘safety lock’) to ‘on’ solely in the presence of the appropriate RNA target substrate. Both proof-of-concept and the mechanism of action of this man-made riboswitch are demonstrated using hepatitis delta virus ribozymes that cleave RNA transcripts derived from the hepatitis B and C viruses. To our knowledge, this is the first report of a ribozyme bearing a target-dependent module that is activated by its RNA substrate, an arrangement which greatly diminishes non-specific effects. This new approach provides a highly specific and improved tool with significant potential for application in the fields of both functional genomics and gene therapy.


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