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Quantitative Comparison of cis-Regulatory Element (CRE) Activities in Transgenic Drosophila melanogaster
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Quantitative Comparison of cis-Regulatory Element (CRE) Activities in Transgenic Drosophila melanogaster

Published on: December 19, 2011

Reduction of Cre recombinase toxicity in proliferating Drosophila cells by estrogen-dependent activity regulation

D Heidmann1, C F Lehner

  • 1Department of Genetics, University of Bayreuth, 95440 Bayreuth, Germany.

Development Genes and Evolution
|October 31, 2001
PubMed

Insights

Researchers developed new Cre recombinase tools for Drosophila melanogaster, overcoming toxicity issues associated with chronic expression. The estrogen-dependent Cre-Estrogen Binding Domain (Cre-EBD) system allows controlled recombination, enhancing genetic manipulation in this model organism.

Area of Science:

  • Molecular Biology
  • Genetics
  • Developmental Biology

Background:

  • The Cre/loxP system is a powerful tool for genome engineering across species.
  • Drosophila melanogaster primarily uses the FLP/FRT system for generating genetic mosaics.
  • Existing Cre/loxP tools in Drosophila have limitations, particularly regarding cell toxicity.

Purpose of the Study:

  • To develop novel Cre recombinase tools for enhanced genetic manipulation in Drosophila melanogaster.
  • To overcome the toxicity associated with constitutive Cre recombinase expression in proliferating cells.
  • To establish an inducible and controllable Cre/loxP system for Drosophila research.

Main Methods:

  • Generation of transgenic Drosophila lines expressing Cre recombinase under the UAS/GAL4 system (UAST-cre).
  • Development of a modified Cre recombinase fused to the estrogen receptor binding domain (UASP-cre-EBD).
  • Assessment of Cre recombinase activity and cellular toxicity in response to estrogen treatment.

Main Results:

  • Constitutive expression of Cre recombinase (UAST-cre) demonstrated toxicity in proliferating Drosophila cells.
  • The Cre-Estrogen Binding Domain (Cre-EBD) fusion protein enabled estrogen-dependent Cre recombinase activity.
  • Estrogen-mediated regulation effectively dissociated Cre recombinase activity from cellular toxicity.

Conclusions:

  • The UASP-cre-EBD system provides a safer and more controllable method for site-specific recombination in Drosophila.
  • This new system expands the genetic toolkit for Drosophila melanogaster, facilitating advanced clonal analysis and genome engineering.
  • Estrogen-inducible Cre recombinase activity offers a significant improvement over constitutive systems for studying gene function in vivo.

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