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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...
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...
Pre-mRNA Processing: RNA Splicing01:32

Pre-mRNA Processing: 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...
Exon Recombination02:32

Exon Recombination

The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes. 
Exon shuffling follows “splice frame rules.” Each exon has three reading...

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Related Experiment Video

Updated: Jun 22, 2026

Synthesis of an Intein-mediated Artificial Protein Hydrogel
15:06

Synthesis of an Intein-mediated Artificial Protein Hydrogel

Published on: January 27, 2014

Traceless protein splicing utilizing evolved split inteins.

Steve W Lockless1, Tom W Muir

  • 1Laboratory of Synthetic Protein Chemistry, The Rockefeller University, New York, NY 10065, USA.

Proceedings of the National Academy of Sciences of the United States of America
|June 23, 2009
PubMed
Summary
This summary is machine-generated.

Split inteins are key to protein splicing and modification. Researchers identified a branched intermediate affecting splicing efficiency and evolved new split inteins for improved protein engineering applications.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Protein Engineering

Background:

  • Split inteins are protein fragments that mediate protein splicing, a process crucial for restoring function in host proteins.
  • They are increasingly utilized in protein engineering for creating modified proteins like cyclical, labeled, and fluorescently tagged variants.
  • Achieving seamless and highly efficient protein splicing is an ongoing goal in the field.

Purpose of the Study:

  • To identify key factors determining split intein splicing efficiency at different junctions.
  • To develop an evolved set of split inteins with altered specificity, higher efficiency, and temperature tolerance.
  • To investigate the mechanisms behind long-range effects of mutations on intein activity.

Main Methods:

  • Experimental identification of the branched intermediate as a determinant of splicing efficiency.
  • Development of a cell-based selection scheme to evolve split inteins.
  • Analysis of mutations in evolved inteins and their positions relative to the active site.

Main Results:

  • The branched intermediate was identified as a critical factor influencing splicing efficiency.
  • Evolved split inteins demonstrated high efficiency at diverse splice junctions and elevated temperatures.
  • Mutations conferring altered specificity were found at sites distant from the intein active site.

Conclusions:

  • Understanding the branched intermediate is crucial for optimizing split intein-mediated splicing.
  • The developed cell-based selection is effective for engineering split inteins with tailored properties.
  • A hypothesis suggests a coevolving amino acid network mediates long-range allosteric effects in split inteins.