Allosteric selection of ribozymes that respond to the second messengers cGMP and cAMP

Makoto Koizumi, Garrett Soukup, J. N Q Kerr, Ronald R. Breaker

Research output: Contribution to journalArticle

151 Scopus citations

Abstract

RNA transcripts containing the hammerhead ribozyme have been engineered to self-destruct in the presence of specific nucleoside 3',5'-cyclic monophosphate compounds. These RNA molecular switches were created by a new combinatorial strategy termed 'allosteric selection,' which favors the emergence of ribozymes that rapidly self-cleave only when incubated with their corresponding effector compounds. Representative RNAs exhibit 5,000- fold activation upon cGMP or cAMP addition, display precise molecular recognition characteristics, and operate with catalytic rates that match those exhibited by unaltered ribozymes. These findings demonstrate that a vast number of ligand-responsive ribozymes with dynamic structural characteristics can be generated in a massively parallel fashion. Moreover, optimized allosteric ribozymes could serve as highly selective sensors of chemical agents or as unique genetic control elements for the programmed destruction of cellular RNAs.

Original languageEnglish
Pages (from-to)1062-1071
Number of pages10
JournalNature Structural Biology
Volume6
Issue number11
DOIs
Publication statusPublished - 1999

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All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Genetics
  • Structural Biology

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