PUBLICATION
A live zebrafish-based screening system for human nuclear receptor ligand and cofactor discovery
- Authors
- Tiefenbach, J., Moll, P.R., Nelson, M.R., Hu, C., Baev, L., Kislinger, T., and Krause, H.M.
- ID
- ZDB-PUB-100330-31
- Date
- 2010
- Source
- PLoS One 5(3): e9797 (Journal)
- Registered Authors
- Hu, Chun, Krause, Henry, Nelson, Meryl, Tiefenbach, Jens
- Keywords
- Embryos, Cofactors (biochemistry), Zebrafish, Epidermis, Eye muscles, Cell fusion, Eyes, Fishes
- MeSH Terms
-
- Animals
- Base Sequence
- Chromatography, Affinity/methods
- Gene Expression Regulation, Developmental
- Genetic Techniques*
- Green Fluorescent Proteins/chemistry
- Hormones
- Humans
- Ligands*
- Molecular Sequence Data
- Plasmids/metabolism
- Protein Isoforms
- Receptors, Cytoplasmic and Nuclear/metabolism*
- Transfection
- Zebrafish
- PubMed
- 20339547 Full text @ PLoS One
Citation
Tiefenbach, J., Moll, P.R., Nelson, M.R., Hu, C., Baev, L., Kislinger, T., and Krause, H.M. (2010) A live zebrafish-based screening system for human nuclear receptor ligand and cofactor discovery. PLoS One. 5(3):e9797.
Abstract
Nuclear receptors (NRs) belong to a superfamily of transcription factors that regulate numerous homeostatic, metabolic and reproductive processes. Taken together with their modulation by small lipophilic molecules, they also represent an important and successful class of drug targets. Although many NRs have been targeted successfully, the majority have not, and one third are still orphans. Here we report the development of an in vivo GFP-based reporter system suitable for monitoring NR activities in all cells and tissues using live zebrafish (Danio rerio). The human NR fusion proteins used also contain a new affinity tag cassette allowing the purification of receptors with bound molecules from responsive tissues. We show that these constructs 1) respond as expected to endogenous zebrafish hormones and cofactors, 2) facilitate efficient receptor and cofactor purification, 3) respond robustly to NR hormones and drugs and 4) yield readily quantifiable signals. Transgenic lines representing the majority of human NRs have been established and are available for the investigation of tissue- and isoform-specific ligands and cofactors.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping