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

Nat Struct Biol. 1999 Nov;6(11):1062-71. doi: 10.1038/14947.


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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Acids / pharmacology
  • Allosteric Regulation / drug effects
  • Allosteric Site / drug effects
  • Base Sequence
  • Catalysis / drug effects
  • Cyclic AMP / pharmacology*
  • Cyclic CMP / pharmacology
  • Cyclic GMP / pharmacology*
  • Enzyme Activation / drug effects
  • Genetic Engineering / methods
  • Kinetics
  • Molecular Sequence Data
  • Nucleic Acid Conformation / drug effects
  • Oligonucleotide Array Sequence Analysis / methods
  • RNA, Catalytic / chemistry*
  • RNA, Catalytic / genetics
  • RNA, Catalytic / metabolism*
  • Structure-Activity Relationship
  • Thermodynamics


  • Acids
  • RNA, Catalytic
  • hammerhead ribozyme
  • Cyclic CMP
  • Cyclic AMP
  • Cyclic GMP