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Germ cell specific promoter drives ectopic transgene expression during embryogenesis
B Bhullar1, J V Schmidt, T Truong
1Department of Biochemistry & Molecular Biology, University of Calgary, Calgary, Alberta, Canada.
Molecular Reproduction and Development
|May 4, 2001
Summary
The phosphoglycerate kinase 2 (Pgk2) promoter drives Cre recombinase expression specifically in adult male germ cells. However, transient, ectopic expression occurs during embryogenesis, potentially impacting Cre-lox studies.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Genetics
Background:
- The phosphoglycerate kinase 2 (Pgk2) promoter is utilized for targeting gene expression in male germ cells.
- Previous studies using Cre-lox systems have reported ectopic Cre expression with various promoters, but its timing and extent remain uncharacterized.
Purpose of the Study:
- To characterize the expression pattern of the Pgk2Cre transgene in mice.
- To assess the suitability of the Pgk2 promoter for Cre-lox based genetic manipulation in male germ cells.
Main Methods:
- Generation of Pgk2Cre transgenic mice.
- Mating of Pgk2Cre mice with H19loxP and lacZ-loxP indicator strains.
- Analysis of Cre-mediated recombination events in adult and embryonic tissues.
- RT-PCR to detect endogenous Pgk2 mRNA expression.
Main Results:
- Pgk2Cre transgene expression is specific to spermatocytes and spermatids in adult mice.
- Recombination events were detected in various embryonic tissues, indicating ectopic Cre expression between days 11 and 15 of embryogenesis.
- Endogenous Pgk2 mRNA was not detected during embryogenesis or in adult non-testis tissues.
Conclusions:
- The Pgk2 promoter is suitable for targeting gene expression to meiotic male germ cells.
- Transient ectopic expression during embryogenesis may affect studies utilizing Cre recombinase with the Pgk2 promoter.