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Updated: Jul 1, 2025

A Cre-Lox P Recombination Approach for the Detection of Cell Fusion In Vivo
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Published on: January 4, 2012

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Glycosylation as a tracer of off-target Cre-lox activation in development

Leandre M Glendenning1, Kalob M Reynero1, Brian A Cobb1

  • 1Department of Pathology, Case Western Reserve University School of Medicine, 10900 Euclid Avenue, Cleveland, OH 44106-7288, United States.

Glycobiology
|March 4, 2024
PubMed

Insights

Plasma membrane glycans can effectively track Cre-lox system activity in stem cells. This method improves the accuracy of lineage-specific genome editing by revealing off-target effects and promoter expression patterns.

Area of Science:

  • Genetics and Genomics
  • Developmental Biology
  • Molecular Biology

Background:

  • The Cre-lox system is a powerful tool for inducible and lineage-specific genome editing in vivo.
  • Challenges include incomplete penetrance and off-target effects from transient Cre expression in stem or precursor cells, complicating data interpretation.
  • Identifying Cre-driver promoter expression across cell lineages is vital for research reproducibility.

Purpose of the Study:

  • To assess the utility of plasma membrane glycans as markers for Cre-lox system activity.
  • To evaluate the specificity and penetrance of Cre-driver promoter expression in vivo.
  • To address limitations in current Cre-lox models, such as the Tie2-Cre system.

Main Methods:

  • Utilized the Tie2 Cre-driver strain as a model system.
  • Investigated plasma membrane glycans as indicators of Cre-mediated recombination.
  • Analyzed Cre-driver promoter expression patterns along cell lineages.

Main Results:

  • Demonstrated that plasma membrane glycans serve as effective markers for Cre-lox system penetrance and specificity.
  • Confirmed transient Cre expression in early precursor cells, impacting lineage models.
  • Highlighted the ubiquitous nature of plasma membrane glycan expression.

Conclusions:

  • Plasma membrane glycans offer a valuable readout for assessing Cre-lox system performance in vivo.
  • This approach enhances the reliability of genome editing studies by detecting off-target effects.
  • The findings improve the rigor and reproducibility of Cre-lox-based research, particularly in developmental and genetic studies.
Keywords:
Cre-loxST6Gal1Tie2glycosylationsialylation

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