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Identification of thyroid hormone response elements in vivo using mice expressing a tagged thyroid hormone receptor alpha 1.


Dudazy-Gralla S, Nordström K, Hofmann PJ, Abdul Meseh D, Schomburg L, Vennström B, Mittag J

Thyroid hormone receptor alpha 1 (TRα1) is well recognized for its importance in brain development. However, due to the difficulties in predicting thyroid hormone response elements (TREs) in silico and the lack of suitable antibodies against TRα1 for chromatin immunoprecipitation, only a few direct TRα1 target genes have been identified in the brain. Here we demonstrate that mice expressing a TRα1-GFP fusion protein from the endogenous TRα locus provide a valuable animal model to identify TRα1 target genes. To this end, we analyzed DNA-TRα1 interactions in vivo using chromatin immunoprecipitation with an anti-GFP antibody. We validated our system using established TREs from neurogranin and hairless, and by verifying additional TREs from known TRα1 target genes in brain and heart. Moreover, our model system enabled the identification of novel TRα1 target genes such as RNF166. Our results demonstrate that transgenic mice expressing a tagged nuclear receptor constitute a feasible approach to study receptor-DNA interactions in vivo, circumventing the need for specific antibodies. Models like the TRα1-GFP mice may thus pave the way for genome-wide mapping of nuclear receptor binding sites, and advance the identification of novel target genes in vivo.

Tags
Bioruptor
Chromatin Shearing
ChIP-qPCR

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Published
February, 2013

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