ORCID: https://orcid.org/0000-0001-8270-1449 und Leonhardt, Heinrich
(2016):
Determination of local chromatin composition by CasID.
In: Nucleus, Vol. 7, No. 5: pp. 476-484
[PDF, 1MB]
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.
Item Type: | Journal article |
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EU Funded Grant Agreement Number: | 638218 |
EU Projects: | Horizon 2020 > ERC Grants > ERC Starting Grant > ERC Grant 638218: MolStruKT - Molecular structure and cell cycle regulated assembly of the kinetochore |
Faculties: | Chemistry and Pharmacy > Department of Biochemistry Chemistry and Pharmacy > GeneCenter |
Subjects: | 500 Science > 540 Chemistry |
URN: | urn:nbn:de:bvb:19-epub-76122-3 |
Language: | English |
Item ID: | 76122 |
Date Deposited: | 01. Jun 2021 09:25 |
Last Modified: | 01. Jun 2021 09:25 |