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Ribozymes02:47

Ribozymes

The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can be...
Ribozymes02:47

Ribozymes

The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can be...
Allosteric Regulation01:08

Allosteric Regulation

Allosteric regulation of enzymes occurs when the binding of an effector molecule to a site that is different from the active site causes a change in the enzymatic activity. This alternate site is called an allosteric site, and an enzyme can contain more than one of these sites. Allosteric regulation can either be positive or negative, resulting in an increase or decrease in enzyme activity. Most enzymes that display allosteric regulation are metabolic enzymes involved in the degradation or...
Allosteric Regulation01:08

Allosteric Regulation

Allosteric regulation of enzymes occurs when the binding of an effector molecule to a site that is different from the active site causes a change in the enzymatic activity. This alternate site is called an allosteric site, and an enzyme can contain more than one of these sites. Allosteric regulation can either be positive or negative, resulting in an increase or decrease in enzyme activity. Most enzymes that display allosteric regulation are metabolic enzymes involved in the degradation or...
Riboswitches01:56

Riboswitches

Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
Allosteric Proteins-ATCase01:19

Allosteric Proteins-ATCase

Binding sites linkages can regulate a protein's function.  For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to  N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis pathway,...

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Related Experiment Video

Updated: Jun 23, 2026

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
09:16

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

In vitro selection of allosteric ribozymes.

Nicolas Piganeau1

  • 1Department of Chemistry, Institute for Biochemistry and Molecular Biology, University of Hamburg, Hamburg, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|April 21, 2009
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel in vitro selection method to create allosteric ribozymes that respond to specific drugs like doxycycline. These self-cleaving ribozymes can be further modified into aptamers for diverse applications.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Synthetic Biology

Background:

  • In vitro selection is a powerful technique for isolating functional nucleic acid sequences.
  • Ribozymes, catalytic RNA molecules, can be engineered for specific functions.
  • Allosteric ribozymes offer precise control over enzymatic activity.

Purpose of the Study:

  • To develop a de novo in vitro selection strategy for allosteric self-cleaving ribozymes.
  • To engineer ribozymes that are specifically inhibited by small molecules (drugs).
  • To demonstrate the selection of hammerhead ribozymes responsive to doxycycline and pefloxacin.

Main Methods:

  • Utilized in vitro selection to screen large libraries for desired ribozyme activity.
  • Applied the selection strategy to isolate hammerhead ribozymes.
  • Tested the selected ribozymes for inhibition by specific drugs at sub-micromolar concentrations.
  • Investigated the conversion of selected ribozymes into aptamers via point mutation.

Main Results:

  • Successfully selected allosteric hammerhead ribozymes.
  • Achieved specific inhibition of selected ribozymes by doxycycline and pefloxacin at sub-micromolar levels.
  • Demonstrated that these ribozymes can be converted into aptamers by altering the catalytic center.

Conclusions:

  • The described in vitro selection method is effective for de novo selection of drug-responsive ribozymes.
  • Selected ribozymes represent a new class of allosteric regulators with potential therapeutic or diagnostic applications.
  • The ability to convert these ribozymes into aptamers broadens their utility in molecular biology and biotechnology.