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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...
DNA-only Transposons02:57

DNA-only Transposons

DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
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...

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Related Experiment Video

Updated: May 25, 2026

Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
09:02

Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

Ribozyme-mediated trans insertion-splicing into target RNAs.

P Patrick Dotson1, Jonathan Hart, Christopher Noe

  • 1Department of Physiology, University of Kentucky, Lexington, KY, USA.

Methods in Molecular Biology (Clifton, N.J.)
|February 9, 2012
PubMed
Summary
This summary is machine-generated.

Trans insertion-splicing (TIS) enables precise RNA editing by inserting donor RNA into acceptor RNA. This method utilizes a specific ribozyme for targeted RNA modification, detailed in system design and protocols.

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Last Updated: May 25, 2026

Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
09:02

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Published on: January 8, 2015

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

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
10:06

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells

Published on: April 26, 2017

Area of Science:

  • Molecular Biology
  • Biochemistry
  • RNA Therapeutics

Background:

  • The trans insertion-splicing (TIS) reaction is a powerful molecular tool.
  • It allows for the site-specific integration of RNA sequences.
  • This technique is crucial for RNA-based research and therapeutic applications.

Purpose of the Study:

  • To describe the trans insertion-splicing (TIS) reaction.
  • To detail the system design for TIS.
  • To outline the ribozyme preparation and reaction protocol.

Main Methods:

  • Utilizing a group I intron-derived ribozyme from Pneumocystis carinii.
  • Designing the RNA donor and acceptor substrates for TIS.
  • Optimizing the reaction conditions and purification methods.

Main Results:

  • Demonstration of site-specific RNA insertion using the TIS system.
  • Successful preparation of the catalytic ribozyme.
  • Establishment of a comprehensive protocol for conducting TIS reactions.

Conclusions:

  • The TIS reaction provides a robust method for targeted RNA manipulation.
  • The described protocol facilitates the application of TIS in various research settings.
  • This technique holds potential for advancing RNA engineering and synthetic biology.