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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...
RNA Structure01:19

RNA Structure

The basic structure of RNA consists of a string of ribonucleotides attached by phosphodiester bonds. Although most RNA is single-stranded, it can form complex secondary and tertiary structures. Such structures play essential roles in the regulation of transcription and translation.
Different Types of RNA Have the Same Basic Structure
There are three main types of ribonucleic acid (RNA) involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). All three...
RNA Structure01:23

RNA Structure

Overview
The basic structure of RNA consists of a five-carbon sugar and one of four nitrogenous bases. Although most RNA is single-stranded, it can form complex secondary and tertiary structures. Such structures play essential roles in the regulation of transcription and translation.
Different Types of RNA Have the Same Basic Structure
There are three main types of ribonucleic acid (RNA): messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). All three RNA types consist of a...
RNA Structure01:23

RNA Structure

Overview
The basic structure of RNA consists of a five-carbon sugar and one of four nitrogenous bases. Although most RNA is single-stranded, it can form complex secondary and tertiary structures. Such structures play essential roles in the regulation of transcription and translation.
Different Types of RNA Have the Same Basic Structure
There are three main types of ribonucleic acid (RNA): messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). All three RNA types consist of a...
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...

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Design and function of triplex hairpin ribozymes.

Guillermo Aquino-Jarquin1, Ramiro Rojas-Hernández, Luis Marat Alvarez-Salas

  • 1Departamento de Fisiología, Biofísica y Neurociencias, CINVESTAV, México D.F, México.

Methods in Molecular Biology (Clifton, N.J.)
|April 14, 2010
PubMed
Summary

Triplex ribozymes enable multiple ribozyme activities for enhanced RNA cleavage. This novel system offers a versatile platform for developing advanced ribozyme-based therapies and technologies.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • RNA Therapeutics

Background:

  • Ribozymes (RZs) are catalytic RNA molecules with therapeutic potential.
  • Tandem expression of multiple ribozymes often leads to reduced individual activity.
  • Novel expression systems are needed to enhance the efficacy of multiple ribozymes.

Purpose of the Study:

  • To develop and characterize a triplex ribozyme system for multiplex RNA expression and cleavage.
  • To engineer triplex expression modules for targeting single or multiple RNA sites.
  • To evaluate the in vitro and in vivo efficacy of the triplex ribozyme system.

Main Methods:

  • Construction of triplex expression modules using hairpin ribozyme cassettes.
  • Generation of multiplex vectors with single or multiple specificity via tandem cloning.
  • In vitro cis- and trans-cleavage assays using radioactive-labeled targets.
  • In vivo assessment in cultured cells using ribonuclease protection assays (RPAs) and RT-PCR.

Main Results:

  • Triplex ribozymes demonstrated enhanced target cleavage compared to tandem-expressed RZs.
  • The system allows for engineering of single or multiple RNA targeting capabilities.
  • Successful validation of triplex ribozyme activity both in vitro and in vivo.

Conclusions:

  • Triplex ribozyme configurations provide a robust platform for multiplex ribozyme expression.
  • This system enhances RNA cleavage efficiency and offers versatile targeting options.
  • The developed triplex ribozyme system lays the groundwork for future ribozyme-based therapies and technologies.