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Related Concept Videos

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...
Enzymes and Activation Energy01:13

Enzymes and Activation Energy

The activation energy (or free energy of activation), abbreviated as Ea, is the small amount of energy input necessary for all chemical reactions to occur. During chemical reactions, certain chemical bonds break, and new ones form. For example, when a glucose molecule breaks down, bonds between the molecule's carbon atoms break. Since these are energy-storing bonds, they release energy when broken. However, the molecule must be somewhat contorted to get into a state that allows the bonds to...
Enzymes and Activation Energy01:13

Enzymes and Activation Energy

The activation energy (or free energy of activation), abbreviated as Ea, is the small amount of energy input necessary for all chemical reactions to occur. During chemical reactions, certain chemical bonds break, and new ones form. For example, when a glucose molecule breaks down, bonds between the molecule's carbon atoms break. Since these are energy-storing bonds, they release energy when broken. However, the molecule must be somewhat contorted to get into a state that allows the bonds to...
ATP Synthase: Mechanism01:48

ATP Synthase: Mechanism

In animals, the mitochondrial F1F0 ATP synthase is the key protein that synthesizes ATP molecules through a complex catalytic mechanism. While the nuclear genome encodes the majority of ATP synthase subunits, the mitochondrial genome encodes some of the enzyme's most critical components. The formation of this multi-subunit enzyme is a complex multi-step process regulated at the level of transcription, translation, and assembly. Defects in one or more of these steps can result in decreased ATP...
ATP Driven Pumps II: P-type Pumps01:34

ATP Driven Pumps II: P-type Pumps

The P-type pumps are a large family of integral membrane transporter ATPases. They are divided into five major types based on substrate specificity, from I to V.
A typical P-type pump has three cytosolic domains: nucleotide-binding (N), phosphorylation (P), and activator (A) domains. These domains are connected to the membrane-spanning helices by short amino acid segments. ATP hydrolysis and covalent phosphoenzyme intermediate formation are crucial parts of the catalytic cycle. At the highly...

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Preparation, Purification, and Use of Fatty Acid-containing Liposomes
10:43

Preparation, Purification, and Use of Fatty Acid-containing Liposomes

Published on: February 9, 2018

Highly active low magnesium hammerhead ribozyme.

Agnieszka Fedoruk-Wyszomirska1, Maciej Szymański, Eliza Wyszko

  • 1Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznań, Poland.

Journal of Biochemistry
|January 7, 2009
PubMed
Summary
This summary is machine-generated.

Hammerhead ribozymes can now inhibit gene expression efficiently in cells. Modified ribozymes work well at low magnesium levels, improving gene silencing strategies.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • RNA Therapeutics

Background:

  • Hammerhead (HH) ribozymes are enzymes that can specifically degrade target mRNA, offering a method for gene expression inhibition.
  • The catalytic efficiency of HH ribozymes is highly sensitive to magnesium ion concentration, with optimal levels not found in cellular environments.
  • Stabilizing tertiary interactions in HH ribozymes can enhance activity at low magnesium concentrations.

Purpose of the Study:

  • To design and validate a novel HH ribozyme capable of efficient catalysis at low magnesium concentrations.
  • To assess the in vitro and cellular efficacy of the designed ribozyme for gene silencing.
  • To compare the performance of the modified ribozyme against existing minimal core ribozymes and DNAzymes.

Main Methods:

  • Design of a HH ribozyme incorporating stabilizing GAAA tetraloop and receptor motifs.
  • In vitro assessment of ribozyme catalytic activity at submillimolar Mg(2+) concentrations.
  • Evaluation of ribozyme performance in cultured cells for gene expression inhibition.
  • Comparison with unmodified and locked nucleic acid (LNA)-modified extended ribozymes and DNAzymes.

Main Results:

  • The designed HH ribozyme efficiently catalyzed target RNA hydrolysis at submillimolar Mg(2+) concentrations in vitro.
  • The ribozyme demonstrated efficacy in cultured cells, indicating successful gene silencing.
  • Both unmodified and LNA-modified extended ribozymes showed superior performance compared to the minimal core ribozyme and DNAzyme.

Conclusions:

  • Stabilization of HH ribozymes through tertiary interactions enables efficient catalysis at physiologically relevant low magnesium concentrations.
  • The developed ribozymes represent a promising tool for gene silencing applications in both in vitro and cellular settings.
  • Modified extended ribozymes offer enhanced performance over minimal ribozymes and DNAzymes for specific mRNA degradation.