L773:0962 8819
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In vitro differentiated adipocytes from a Foxc2 reporter knock-in mouse as screening tool.
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- Cederberg, Anna, 1972 (author)
- Gothenburg University,Göteborgs universitet,Institutionen för biomedicin, avdelningen för medicinsk genetik och klinisk genetik,Institute of Biomedicine, Department of Medical and Clinical Genetics
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- Grände, Mats, 1975 (author)
- Gothenburg University,Göteborgs universitet,Institutionen för biomedicin, avdelningen för medicinsk genetik och klinisk genetik,Institute of Biomedicine, Department of Medical and Clinical Genetics
- Rhedin, Magdalena (author)
- show more...
- Peng, Xiao-Rong (author)
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- (creator_code:org_t)
- 2009-05-28
- 2009
- English.
- In: Transgenic research. - : Springer Science and Business Media LLC. - 1573-9368 .- 0962-8819. ; 18:6, s. 889-97
- Journal article (peer-reviewed)
Abstract
Subject headings
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- We have developed a generic model for in vitro high-throughput screening for agents regulating transcription of genes in the mouse genome here exemplified by Foxc2, a forkhead transcription factor involved in regulation of adipocyte metabolism. We made a Foxc2-LacZ reporter "knock-in" mouse in which one of the two Foxc2 alleles has been inactivated and replaced by a LacZ reporter gene. Mouse embryonic fibroblasts, derived from such mice, were differentiated in vitro to adipocytes and used in cell-based screens. Forskolin as well as 12-O-tetradecanoylphorbol-13-acetate (TPA) increased levels of Foxc2nLacZ fusion protein. We could also demonstrate that this was paralleled by an increase in Foxc2 mRNA, transcribed from the wild type allele. This generic method offers a novel way of identifying both positive and negative upstream regulators of a gene, using high-throughput screening methodology. In a cell-based screen using such methodology we demonstrate efficacy by identifying NKH477 as a Foxc2 activating compound.
Keyword
- Adipocytes
- cytology
- metabolism
- Animals
- Cell Differentiation
- drug effects
- Cells
- Cultured
- Drug Evaluation
- Preclinical
- Embryo
- Mammalian
- cytology
- Female
- Fibroblasts
- cytology
- metabolism
- Forkhead Transcription Factors
- genetics
- Gene Knock-In Techniques
- Lac Operon
- Male
- Mice
- Small Molecule Libraries
Publication and Content Type
- ref (subject category)
- art (subject category)
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