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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.
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Rapidly dividing tumors, embryos, and wounded tissues require more oxygen than usual, lowering the oxygen concentration in the blood. At low oxygen or hypoxic conditions, an oxygen-sensitive transcription factor called the hypoxia-inducible factor 1 or HIF1 is activated. HIF1 is a dimeric protein of alpha (ɑ) and beta (β) subunits.  Under optimal oxygen conditions, HIF1β is present in the nucleus while HIF1ɑ remains in the cytosol. HIF1ɑ is hydroxylated by prolyl hydroxylase and factor...

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Related Experiment Video

Updated: Jun 23, 2026

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium
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Published on: May 6, 2018

Conditional gene targeting in mouse high endothelial venules.

Hiroto Kawashima1, Jotaro Hirakawa, Yuki Tobisawa

  • 1Laboratory of Microbiology and Immunology, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka, Japan. kawashih@ushizuoka-ken.ac.jp

Journal of Immunology (Baltimore, Md. : 1950)
|April 22, 2009
PubMed
Summary

A new transgenic mouse line specifically expresses Cre recombinase in high endothelial venules (HEVs) and colon, aiding research into tissue-specific gene functions. This tool is valuable for studying immune responses and gene regulation in various organs.

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Area of Science:

  • Immunology
  • Developmental Biology
  • Genetics

Background:

  • High endothelial venules (HEVs) are specialized blood vessels crucial for lymphocyte homing in secondary lymphoid organs.
  • Understanding HEV-specific gene expression is vital for dissecting immune responses and lymph node development.
  • Existing tools lack specificity for targeting HEVs, limiting functional studies.

Purpose of the Study:

  • To develop a novel transgenic mouse model for studying gene function specifically in HEVs.
  • To characterize the expression pattern of N-acetylglucosamine-6-O-sulfotransferase 2 (GlcNAc6ST-2) in vivo.
  • To establish a tool for Cre/loxP-mediated gene manipulation in HEVs and other GlcNAc6ST-2-expressing tissues.

Main Methods:

  • Generated a transgenic mouse line using bacterial artificial chromosome recombineering to drive Cre recombinase expression under the GlcNAc6ST-2 promoter.
  • Crossed transgenic mice with ROSA26 reporter mice to visualize Cre activity via lacZ expression.
  • Analyzed Cre expression in secondary lymphoid organs, colon, and other tissues using immunohistochemistry and RT-PCR.

Main Results:

  • Cre recombinase was specifically expressed in HEVs of secondary lymphoid organs, correlating with HEV maturation.
  • Strong Cre expression was observed in colonic villi, regulated by commensal bacteria.
  • Off-target expression was detected in a small subset of cells in the brain, testis, stomach, small intestine, and lung.

Conclusions:

  • The novel GlcNAc6ST-2-driven Cre mouse line accurately reports HEV-specific gene expression in lymphoid organs.
  • This model also captures bacterial-regulated gene expression in the colon, offering a dual-purpose research tool.
  • The restricted expression pattern makes this transgenic line valuable for tissue-specific gene function analysis using the Cre/loxP system.