PUBLICATION
Application of the dual-luciferase reporter assay to the analysis of promoter activity in zebrafish embryos
- Authors
- Alcaraz-Perez, F., Mulero, V., and Cayuela, M.L.
- ID
- ZDB-PUB-081029-26
- Date
- 2008
- Source
- BMC Biotechnology 8: 81 (Journal)
- Registered Authors
- Mulero, Victor
- Keywords
- none
- MeSH Terms
-
- Animals
- Embryonic Development
- Gene Knockdown Techniques
- Genes, Reporter*
- Luciferases, Firefly/genetics*
- Luciferases, Firefly/metabolism
- Luciferases, Renilla/genetics*
- Luciferases, Renilla/metabolism
- NF-kappa B/metabolism
- Promoter Regions, Genetic*
- Two-Hybrid System Techniques
- Zebrafish/embryology
- Zebrafish/genetics*
- Zebrafish/metabolism
- PubMed
- 18954456 Full text @ BMC Biotechnol.
Citation
Alcaraz-Perez, F., Mulero, V., and Cayuela, M.L. (2008) Application of the dual-luciferase reporter assay to the analysis of promoter activity in zebrafish embryos. BMC Biotechnology. 8:81.
Abstract
BACKGROUND: The dual-luciferase assay has been widely used in cell lines to determine rapidly but accurately the activity of a given promoter. Although this strategy has proved very useful, it does not allow the promoter and gene function to be analyzed in the context of the whole organism. RESULTS: Here, we present a rapid and sensitive assay based on the classical dual-luciferase reporter technique which can be used as a new tool to characterize the minimum promoter region of a gene as well as the in vivo response of inducible promoters to different stimuli. We illustrate the usefulness of this system for studying both constitutive (telomerase) and inducible (NF-kappaB-dependent) promoters. The flexibility of this assay is demonstrated by induction of the NF-kappaB-dependent promoters using simultaneous microinjection of different pathogen-associated molecular patterns as well as with the use of morpholino-gene mediated knockdown. CONCLUSIONS: This assay has several advantages compared with the classical in vitro (cell lines) and in vivo (transgenic mice) approaches. Among others, the assay allows a rapid and quantitative measurement of the effects of particular genes or drugs in a given promoter in the context of a whole organism and it can also be used in high throughput screening experiments.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping