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Delta ribozyme benefits from a good stability in vitro that becomes outstanding in vivo

Published online by Cambridge University Press:  24 April 2002

DOMINIQUE LÉVESQUE
Affiliation:
Département de biochimie, Université de Sherbrooke, Sherbrooke, Québec, J1H 5N4, Canada
SANAA CHOUFANI
Affiliation:
Département d'anatomie et biologie cellulaire, Université de Sherbrooke, Sherbrooke, Québec, J1H 5N4, Canada
JEAN-PIERRE PERREAULT
Affiliation:
Département de biochimie, Université de Sherbrooke, Sherbrooke, Québec, J1H 5N4, Canada
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Abstract

The stability of a trans-acting delta ribozyme was studied under various conditions. Although in vitro (i.e., in the presence of protein extracts) this delta ribozyme appears to be only slightly more stable than a hammerhead ribozyme, in vivo (i.e., after cell transfection) it exhibits an outstanding stability that manifests itself in the calculated half-life of over 100 h regardless of the means of transfection. The P2 stem, which includes both the 5′ and 3′ ends, is shown to play a critical role in this stability. Direct mutagenesis of the most nuclease susceptible nucleotides failed to generate a more stable ribozyme that retained the same catalytic potential. Clearly, delta ribozyme appears to be well adapted to the human cell environment, and is therefore ideal for the development of a gene-inactivation system.

Type
Research Article
Copyright
2002 RNA Society

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