Journal
NUCLEUS
Volume 7, Issue 5, Pages 476-484Publisher
TAYLOR & FRANCIS INC
DOI: 10.1080/19491034.2016.1239000
Keywords
biotinylation; CRISPR; Cas; CasID; chromatin composition; repetitive elements
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Funding
- Deutsche Forschungsgemeinschaft [SFB1064, SFB646]
- European Research Council [638218]
- European Research Council (ERC) [638218] Funding Source: European Research Council (ERC)
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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.
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