Abstract

To establish a genetic tool for conditional deletion or expression of gene in neurons in a temporally controlled manner, we generated a transgenic mouse (NSE-MerCreMer), which expressed a tamoxifen inducible type of Cre recombinase specifically in neurons. The tamoxifen inducible Cre recombinase (MerCreMer) is a fusion protein containing Cre recombinase with two modified estrogen receptor ligand binding domains at both ends, and is driven by the neural-specific rat neural specific enolase (NSE) promoter. A total of two transgenic lines were established, and expression of MerCreMer in neurons of the central and enteric nervous systems was confirmed. Transcript of MerCreMer was detected in several non-neural tissues such as heart, liver, and kidney in these lines. In the background of the Cre reporter mouse strain Rosa26R, Cre recombinase activity was inducible in neurons of adult NSE-MerCreMer mice treated with tamoxifen by intragastric gavage, but not in those fed with corn oil only. We conclude that NSE-MerCreMer lines will be useful for studying gene functions in neurons for the conditions that Cre-mediated recombination resulting in embryonic lethality, which precludes investigation of gene functions in neurons through later stages of development and in adult.

Highlights

  • Conventional gene knockout in mice can result in early lethality, which prevents study of gene functions through later stages of development and in adult

  • Cre recombinase is a Type I topoisomerase from bacteriophage P1 that catalyzes the site-specific recombination of DNA between loxP elements in both bacteriophage and mammalian cells [1,2,3]

  • Establishment of neural specific enolase (NSE)-MerCreMer Transgenic Mice Transgenic construct NSE-MerCreMer was constructed by linking the rat NSE promoter 59 upstream of the cDNA encoding the tamoxifen inducible Cre recombinase (MerCreMer), which is composed of Cre recombinase and mutated ligand-binding domain (LBD) of the mouse estrogen receptor a (ERa) (Mer) on both ends (Figure 1)

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Summary

Introduction

Conventional gene knockout in mice can result in early lethality, which prevents study of gene functions through later stages of development and in adult. We cloned the rat NSE promoter 59 to the cDNA encoding the MerCreMer and generated transgenic mouse lines that expressed tamoxifen inducible Cre activity in neurons.

Results
Conclusion
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