Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

RNA Splicing01:32

RNA Splicing

Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
RNA Splicing01:32

RNA Splicing

Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
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...
Alternative RNA Splicing02:18

Alternative RNA Splicing

Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
Alternative RNA Splicing02:18

Alternative RNA Splicing

Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

A GTP synthase ribozyme with increased GTP turnover.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same author

The Exceptional Solubility of Cyclic Trimetaphosphate in the Presence of Mg<sup>2+</sup> and Ca<sup>2</sup>.

Life (Basel, Switzerland)·2026
Same author

Repair of Mutated <i>NF1</i> mRNA with Trans-Splicing Group I Intron Ribozymes.

Cancers·2025
Same author

Assembly of catalytic complexes from randomized oligonucleotides.

Science advances·2025
Same author

Principles of <i>in vitro</i> selection of ribozymes from random sequence libraries.

Journal of the Royal Society, Interface·2025
Same author

Weak effects of prebiotically plausible peptides on self-triphosphorylation ribozyme function.

RSC chemical biology·2024

Related Experiment Video

Updated: May 25, 2026

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression
08:54

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression

Published on: March 29, 2019

An in vivo selection method to optimize trans-splicing ribozymes.

Karen E Olson1, Ulrich F Müller

  • 1Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, California 92093, USA.

RNA (New York, N.Y.)
|January 26, 2012
PubMed
Summary

This study introduces a novel in vivo selection method to identify highly efficient group I intron ribozymes for therapeutic trans-splicing. The new technique rapidly screens millions of variants, significantly improving ribozyme design for genetic disorder treatments.

More Related Videos

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
08:53

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

Published on: September 15, 2021

A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA
13:00

A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA

Published on: December 2, 2009

Related Experiment Videos

Last Updated: May 25, 2026

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression
08:54

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression

Published on: March 29, 2019

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
08:53

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

Published on: September 15, 2021

A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA
13:00

A Rapid High-throughput Method for Mapping Ribonucleoproteins (RNPs) on Human pre-mRNA

Published on: December 2, 2009

Area of Science:

  • Molecular Biology
  • Biochemistry
  • RNA Therapeutics

Background:

  • Group I intron ribozymes perform mRNA trans-splicing, offering potential for genetic disorder therapy and targeted cell killing.
  • Current limitations in therapeutic applications stem from low in vivo trans-splicing efficiency.
  • Previous optimization efforts focused on individual ribozyme constructs, limiting scalability.

Purpose of the Study:

  • To develop and validate a high-throughput in vivo selection method for identifying efficient trans-splicing group I intron ribozymes.
  • To overcome the bottleneck of low in vivo efficiency in ribozyme-based therapeutics.
  • To establish a platform for optimizing ribozyme sequences in diverse contexts.

Main Methods:

  • An in vivo antibiotic resistance gene rescue assay was employed.
  • A library of 9 × 10⁶ randomized Tetrahymena thermophila group I intron variants was screened.
  • Bacterial cells expressing functional ribozymes survived chloramphenicol selection.

Main Results:

  • The selection method successfully identified ribozyme variants with significantly enhanced trans-splicing efficiency.
  • This approach facilitates the discovery of optimized ribozymes for specific sequence requirements.
  • The method demonstrates scalability for screening large ribozyme libraries.

Conclusions:

  • A powerful in vivo selection strategy has been established for optimizing group I intron ribozymes.
  • This method accelerates the development of efficient ribozymes for therapeutic trans-splicing applications.
  • The platform is adaptable for optimizing any trans-splicing ribozyme sequence.