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Regulation of Ribozyme Cleavage Activity by Oligonucleotides

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Part of the Methods in Molecular Biology™ book series (MIMB, volume 252)

Abstract

Here, we describe allosteric ribozymes, which are activated by the addition of a short regulator oligonucleotide. The allosteric hammerhead ribozyme, which contains a single-stranded loop instead of stem II, exhibited minimal cleavage of the target RNA; however, it becomes active by the addition of oligonucleotides that bind to the ribozyme. We have also carried out in vitro selection to obtain an allosteric hairpin ribozyme, which is activated with a short oligonucleotide. The ribozyme contains the characteristic hairpin loop with the structure that changes upon regulator oligonucleotide binding. Since both regulations are sequence-specific, ribozymes containing different sequences can be positively and independently controlled by a specific oligonucleotide. Furthermore, these allosteric hairpin loops have the ability to control the activities of other functional RNAs.

Keywords

Ammonium Persulfate Stop Solution Cleavage Activity Cleavage Reaction Hairpin Loop 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Copyright information

© Humana Press Inc., Totowa, NJ 2004

Authors and Affiliations

  1. 1.DNA Chip Reseach Inc.YokohamaJapan
  2. 2.National Institute of Advanced Industrial Science and TechnologySapporoJapan

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