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Here, we show that transcription factors bound to regulatory sequences can be identified by purifying these unique sequences directly from mammalian cells in vivo. Using targeted chromatin purification (TChP), a double-pull-down strategy with a tetracycline-sensitive "hook" bound to a specific promoter, we identify transcription factors bound to the repressed γ-globin gene-associated regulatory regions. After validation of the binding, we show that, in human primary erythroid cells, knockdown of a number of these transcription factors induces γ-globin gene expression. Reactivation of γ-globin gene expression ameliorates the symptoms of β-thalassemia and sickle cell disease, and these factors provide potential targets for the development of therapeutics for treating these patients.

Original publication

DOI

10.1016/j.celrep.2013.07.004

Type

Journal article

Journal

Cell Rep

Publication Date

15/08/2013

Volume

4

Pages

589 - 600

Keywords

Animals, Cells, Cultured, Chromatin, Gene Knockdown Techniques, Humans, Mass Spectrometry, Mice, Mice, Transgenic, Promoter Regions, Genetic, Proteomics, Transcription, Genetic, beta-Globins