Locus-Specific Proteomics by TChP: Targeted Chromatin Purification

• DOI: 10.1016/j.celrep.2013.07.004
• Publication Date: 2013-08-01T20:01:46Z
• Authors: Farzin Pourfarzad; Ali Aghajanirefah; Ernie de Boer; Sara Ten Have; Thamar Bryn van Dijk; Sima Kheradmandkia; Ralph Stadhouders; Supat Thongjuea; Eric Soler; Nynke Gillemans; Marieke von Lindern; Jeroen Demmers; Sjaak Philipsen; Frank Grosveld
• Keywords: Transcription

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