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A live zebrafish-based screening system for human nuclear receptor ligand and cofactor discovery.
- Source :
-
PloS one [PLoS One] 2010 Mar 22; Vol. 5 (3), pp. e9797. Date of Electronic Publication: 2010 Mar 22. - Publication Year :
- 2010
-
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.
- Subjects :
- Animals
Base Sequence
Chromatography, Affinity methods
Gene Expression Regulation, Developmental
Green Fluorescent Proteins chemistry
Hormones
Humans
Molecular Sequence Data
Plasmids metabolism
Protein Isoforms
Transfection
Zebrafish
Genetic Techniques
Ligands
Receptors, Cytoplasmic and Nuclear metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 5
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 20339547
- Full Text :
- https://doi.org/10.1371/journal.pone.0009797