title: Locus-specific proteomics by TChP: targeted chromatin purification.
abstract: 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.
authors.affiliation: Department of Cell Biology, Erasmus MC, Dr. Molewaterplein 50, 3015GE Rotterdam, the Netherlands; Center for Biomedical Genetics, Erasmus MC, Dr. Molewaterplein 50, 3015GE Rotterdam, the Netherlands.
authors.forename: Farzin
authors.initials: F
authors.lastname: Pourfarzad
authors.forename: Ali
authors.initials: A
authors.lastname: Aghajanirefah
authors.forename: Ernie
authors.initials: E
authors.lastname: de Boer
authors.forename: Sara
authors.initials: S
authors.lastname: Ten Have
authors.forename: Thamar
authors.initials: T
authors.lastname: Bryn van Dijk
authors.forename: Sima
authors.initials: S
authors.lastname: Kheradmandkia
authors.forename: Ralph
authors.initials: R
authors.lastname: Stadhouders
authors.forename: Supat
authors.initials: S
authors.lastname: Thongjuea
authors.forename: Eric
authors.initials: E
authors.lastname: Soler
authors.forename: Nynke
authors.initials: N
authors.lastname: Gillemans
authors.forename: Marieke
authors.initials: M
authors.lastname: von Lindern
authors.forename: Jeroen
authors.initials: J
authors.lastname: Demmers
authors.forename: Sjaak
authors.initials: S
authors.lastname: Philipsen
authors.forename: Frank
authors.initials: F
authors.lastname: Grosveld
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002843:Chromatin / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D055785:Gene Knockdown Techniques / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic; D040901:Proteomics / Q000379:methods; D014158:Transcription, Genetic; D055544:beta-Globins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D002843:Chromatin; D055544:beta-Globins