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A live zebrafish-based screening system for human nuclear receptor ligand and cofactor discovery.
Tiefenbach, Jens; Moll, Pamela R; Nelson, Meryl R; Hu, Chun; Baev, Lilia; Kislinger, Thomas; Krause, Henry M.
Afiliação
  • Tiefenbach J; Banting and Best Department of Medical Research, The Terrence Donnelly Centre for Cellular and Biomolecular Research (CCBR), University of Toronto, Toronto, Ontario, Canada.
PLoS One ; 5(3): e9797, 2010 Mar 22.
Article em En | MEDLINE | ID: mdl-20339547
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.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Técnicas Genéticas / Receptores Citoplasmáticos e Nucleares / Ligantes Tipo de estudo: Diagnostic_studies / Screening_studies Limite: Animals / Humans Idioma: En Ano de publicação: 2010 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Técnicas Genéticas / Receptores Citoplasmáticos e Nucleares / Ligantes Tipo de estudo: Diagnostic_studies / Screening_studies Limite: Animals / Humans Idioma: En Ano de publicação: 2010 Tipo de documento: Article