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A Versatile ΦC31 Based Reporter System for Measuring AP-1 and Nrf2 Signaling in Drosophila and in Tissue Culture
- Source :
- PLoS ONE, Vol 7, Iss 4, p e34063 (2012), PLoS ONE
- Publication Year :
- 2012
- Publisher :
- Public Library of Science (PLoS), 2012.
-
Abstract
- This paper describes the construction and characterization of a system of transcriptional reporter genes for monitoring the activity of signaling pathways and gene regulation mechanisms in intact Drosophila, dissected tissues or cultured cells. Transgenic integration of the reporters into the Drosophila germline was performed in a site-directed manner, using ΦC31 integrase. This strategy avoids variable position effects and assures low base level activity and high signal responsiveness. Defined integration sites furthermore enable the experimenter to compare the activity of different reporters in one organism. The reporter constructs have a modular design to facilitate the combination of promoter elements (synthetic transcription factor binding sites or natural regulatory sequences), reporter genes (eGFP, or DsRed.T4), and genomic integration sites. The system was used to analyze and compare the activity and signal response profiles of two stress inducible transcription factors, AP-1 and Nrf2. To complement the transgenic reporter fly lines, tissue culture assays were developed in which the same synthetic ARE and TRE elements control the expression of firefly luciferase.
- Subjects :
- Aging
Anatomy and Physiology
Transcription, Genetic
Cell Culture Techniques
lcsh:Medicine
Tissue Culture Techniques
0302 clinical medicine
Genes, Reporter
Molecular Cell Biology
Homeostasis
Signaling in Cellular Processes
Drosophila Proteins
Bacteriophages
lcsh:Science
Cellular Stress Responses
Genetics
Regulation of gene expression
0303 health sciences
Multidisciplinary
Drosophila Melanogaster
Gene Transfer Techniques
Animal Models
Cell biology
Regulatory sequence
Bioreporter
Genetic Engineering
Drosophila Protein
Research Article
Biotechnology
Signal Transduction
NF-E2-Related Factor 2
DNA transcription
Biology
Molecular Genetics
03 medical and health sciences
Model Organisms
Stress, Physiological
Animals
Gene Regulation
Luciferase
Transcription factor
030304 developmental biology
Reporter gene
Binding Sites
lcsh:R
JNK Mitogen-Activated Protein Kinases
Transcription Factor AP-1
DNA binding site
Gene Expression Regulation
lcsh:Q
Gene expression
Transcriptional Signaling
Physiological Processes
030217 neurology & neurosurgery
Transgenics
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 7
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....c5a74569b60d00f85451b55bd58560ce