Determination of local chromatin composition by CasID. (2nd September 2016)

• Record Type: Journal Article
• Authors: Schmidtmann, Elisabeth; Anton, Tobias; Rombaut, Pascaline; Herzog, Franz; Leonhardt, Heinrich
• Is Part Of: Nucleus. Volume 7:Number 5(2016)
• Page Start: 476
• Page End: 484
• Publication Date: 2016-09-02
• Subjects: biotinylation -- CRISPR/Cas -- CasID -- chromatin composition -- repetitive elements; Cell nuclei -- Periodicals; Biology -- Periodicals; Biology; Cell nuclei; Periodicals; 571.6605
• DOI: 10.1080/19491034.2016.1239000
• Languages: English
• ISSNs: 1949-1034

Abstract

ABSTRACT: Chromatin structure and function are determined by a plethora of proteins whose genome-wide distribution is typically assessed by immunoprecipitation (ChIP). Here, we developed a novel tool to investigate the local chromatin environment at specific DNA sequences. We combined the programmable DNA binding of dCas9 with the promiscuous biotin ligase BirA* (CasID) to biotinylate proteins in the direct vicinity of specific loci. Subsequent streptavidin-mediated precipitation and mass spectrometry identified both known and previously unknown chromatin factors associated with repetitive telomeric, major satellite and minor satellite DNA. With super-resolution microscopy, we confirmed the localization of the putative transcription factor ZNF512 at chromocenters. The versatility of CasID facilitates the systematic elucidation of functional protein complexes and locus-specific chromatin composition.