Related Experiment Video

Updated: Jul 6, 2026

Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis
13:31

Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis

Published on: October 31, 2014

Transposon-mediated mutagenesis in somatic cells: identification of transposon-genomic DNA junctions

David A Largaespada1, Lara S Collier

  • 1Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN.

Methods in Molecular Biology (Clifton, N.J.)
|March 29, 2008
PubMed

Insights

The Sleeping Beauty transposon system aids cancer gene discovery in mouse models. This method uses transposons for insertional mutagenesis and tagging, facilitating the identification of cancer-driving genes.

Area of Science:

  • * Genetics
  • * Cancer Biology
  • * Molecular Biology

Background:

  • * Understanding the genetic basis of tumor formation is vital for effective cancer treatment.
  • * Forward genetic screens with insertional mutagenesis have historically identified key cancer genes in mouse models.
  • * Retroviruses were traditionally used, leading to discoveries in leukemia, lymphoma, and mammary cancer.

Purpose of the Study:

  • * To explore the utility of the Sleeping Beauty transposon system for cancer gene discovery.
  • * To present a polymerase chain reaction-based method for identifying transposon-tagged sequences in tumors.

Main Methods:

  • * Utilizing the Sleeping Beauty transposon system for insertional mutagenesis in mouse models.
  • * Employing the transposon as a molecular tag for efficient gene identification.
  • * Developing and presenting a polymerase chain reaction (PCR)-based cloning technique.

Main Results:

  • * The Sleeping Beauty transposon system enables insertional mutagenesis in the mouse soma.
  • * Transposon insertion facilitates the identification of cancer genes.
  • * The presented PCR method allows for the cloning of transposon-tagged sequences from tumor samples.

Conclusions:

  • * The Sleeping Beauty transposon system is a valuable tool for cancer gene discovery in mouse models.
  • * This system aids in identifying oncogenes and tumor suppressor genes critical for cancer development.
  • * The described PCR-based method streamlines the process of candidate cancer gene identification.

More Related Videos

Determination of the Optimal Chromosomal Location(s) for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach11:12

Determination of the Optimal Chromosomal Location(s) for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach

Published on: September 11, 2017

Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing08:19

Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing

Published on: July 7, 2020

Related Concept Videos

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...
Transposons01:24

Transposons

Transposons, or "jumping genes," are small mobile genetic elements (MGEs) that range from 700 to 40,000 base pairs in length. They are found in all organisms and can move within the same chromosome or transfer to different chromosomes. In some cases, transposons can also jump between different host DNA molecules, such as plasmids or viruses, contributing to genetic variability.Barbara McClintock first discovered these mobile genetic elements in the 1940s while studying maize genetics, and she...
Overview of Transposition and Recombination02:13

Overview of Transposition and Recombination

Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
Non-LTR Retrotransposons03:18

Non-LTR Retrotransposons

As the name suggests, non-LTR retrotransposons lack the long terminal repeats characteristic of the LTR retrotransposons. Additionally, both LTR and non-LTR retrotransposons use distinct mechanisms of mobilization. Non-LTR retrotransposons are further divided into two classes - Long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs), both of which occur abundantly in most mammals, including humans. Some of the active non-LTR retrotransposons in humans are L1...
Gene Conversion02:08

Gene Conversion

Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...