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Related Concept Videos

In-vitro Mutagenesis01:16

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
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...

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Related Experiment Video

Updated: May 10, 2026

Mouse Genome Engineering Using Designer Nucleases
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Published on: April 2, 2014

Controlled insertional mutagenesis using a LINE-1 (ORFeus) gene-trap mouse model.

Kathryn A O'Donnell1, Wenfeng An, Christina T Schrum

  • 1Department of Molecular Biology and Genetics, and High Throughput Biology Center, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. kathryn.odonnell@UTSouthwestern.edu

Proceedings of the National Academy of Sciences of the United States of America
|July 3, 2013
PubMed
Summary

A novel tetracycline-regulated retrotransposon (ORFeus) enables potent, controllable gene mutations in mice. This system shows promise for creating versatile mouse models for genetic research and drug discovery.

Keywords:
L1 retrotransposonmus musculustet-promoterwhite-spotted phenotype

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Area of Science:

  • Genetics
  • Molecular Biology
  • Genomics

Background:

  • Long interspersed element 1 (LINE-1) retrotransposons are mobile genetic elements.
  • Optimized LINE-1 elements can exhibit enhanced retrotransposition activity.
  • Developing controllable genetic tools is crucial for disease modeling.

Purpose of the Study:

  • To create a tetracycline-regulated retrotransposon-based mouse model for inducible and potent mutagenesis.
  • To assess the mutagenic capacity and functionality of the ORFeus element in vivo.
  • To explore the utility of this system as an alternative to DNA transposon-based mutagenesis.

Main Methods:

  • Generation of a tetracycline-inducible ORFeus retrotransposon with a gene-trap cassette.
  • Administration of doxycycline to induce transgene expression and retrotransposition in mice.
  • Phenotypic analysis of resulting mutations, including coat color changes in skin.
  • Breeding with different tetracycline-transactivator lines to assess functional versatility.

Main Results:

  • The tetracycline-regulated ORFeus element (tet-ORFeus) demonstrated robust retrotransposition in somatic tissues upon doxycycline treatment.
  • High-level doxycycline induction during embryogenesis led to a significant mutagenic burden, reducing double transgenic animal numbers.
  • Induction in skin resulted in a white spotting phenotype, indicating somatic mutations affecting melanocyte development.
  • The tet-ORFeus system exhibited dose-dependent, tissue-specific, and temporal-specific mutagenesis potential.

Conclusions:

  • The tet-ORFeus system provides a powerful and regulatable tool for retrotransposon-mediated mutagenesis in mice.
  • This system offers a viable alternative to existing DNA transposon-based mutagenesis strategies.
  • The demonstrated versatility supports its application in generating diverse mouse models for genetic studies.