Related Experiment Video

Updated: Jul 11, 2026

Defining Gene Functions in Tumorigenesis by Ex vivo Ablation of Floxed Alleles in Malignant Peripheral Nerve Sheath Tumor Cells
09:37

Defining Gene Functions in Tumorigenesis by Ex vivo Ablation of Floxed Alleles in Malignant Peripheral Nerve Sheath Tumor Cells

Published on: August 25, 2021

Variable expression of Cre recombinase transgenes precludes reliable prediction of tissue-specific gene disruption by

Tim J Schulz1, Markus Glaubitz, Doreen Kuhlow

  • 1University of Jena, Institute of Nutrition, Jena, Germany.

Plos One
|October 11, 2007
PubMed

Insights

Cre recombinase expression can be lost across generations in transgenic mice, leading to failed gene disruption. Genotyping alone may not confirm gene knockout efficacy, necessitating post hoc analysis.

Area of Science:

  • Genetics
  • Molecular Biology
  • Transgenic Technology

Background:

  • The Cre/loxP system is widely used for generating conditional knockout mouse models for tissue-specific gene disruption.
  • Ensuring reliable Cre recombinase activity is crucial for the accurate generation of these models.

Purpose of the Study:

  • To investigate the phenomenon of declining Cre recombinase expression in a transgenic mouse strain over successive generations.
  • To determine the impact of diminished Cre expression on the efficacy of target gene disruption.

Main Methods:

  • Utilized a transgenic mouse strain engineered for Cre/loxP-mediated gene disruption.
  • Monitored Cre recombinase expression across multiple generations.
  • Assessed target gene disruption at both the genomic and protein expression levels.

Main Results:

  • Observed a progressive loss of Cre recombinase expression in the transgenic strain with increasing generations.
  • Complete abrogation of Cre gene expression was detected at the molecular level, despite intact genomic DNA.
  • This loss of Cre expression resulted in incomplete or absent disruption of the target gene.

Conclusions:

  • Cre recombinase expression can be epigenetically silenced or otherwise lost in transgenic lines over time.
  • Genomic analysis of Cre transgenes may not reliably predict functional gene disruption.
  • Post hoc validation of target gene disruption is essential for all subjects in Cre/loxP studies to ensure experimental validity.

More Related Videos

The Drosophila Imaginal Disc Tumor Model: Visualization and Quantification of Gene Expression and Tumor Invasiveness Using Genetic Mosaics10:31

The Drosophila Imaginal Disc Tumor Model: Visualization and Quantification of Gene Expression and Tumor Invasiveness Using Genetic Mosaics

Published on: October 6, 2016

Conditional Genetic Transsynaptic Tracing in the Embryonic Mouse Brain11:03

Conditional Genetic Transsynaptic Tracing in the Embryonic Mouse Brain

Published on: December 22, 2014

Related Concept Videos

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...
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.
Reporter Genes02:11

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...