ArticleCurrent protocols in molecular biology2015
MARCC (Matrix-Assisted Reader Chromatin Capture): An Antibody-Free Method to Enrich and Analyze Combinatorial Nucleosome Modifications.
Article in Current protocols in molecular biology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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4 citing papers in PubMed.
- Design of genetically encoded sensors to detect nucleosome ubiquitination in live cells.The Journal of cell biology · 2021Article
- Engineered Reader Proteins for Enhanced Detection of Methylated Lysine on Histones.ACS chemical biology · 2020Article
- EBS is a bivalent histone reader that regulates floral phase transition in Arabidopsis.Nature genetics · 2018Article
- General method for rapid purification of native chromatin fragments.The Journal of biological chemistry · 2018Article
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2 authors.
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Abstract
Combinatorial patterns of histone modifications are key indicators of different chromatin states. Most of the current approaches rely on the usage of antibodies to analyze combinatorial histone modifications. Here we detail an antibody-free method named MARCC (Matrix-Assisted Reader Chromatin Capture) to enrich combinatorial histone modifications. The combinatorial patterns are enriched on native nucleosomes extracted from cultured mammalian cells and prepared by micrococcal nuclease digestion. Such enrichment is achieved by recombinant chromatin-interacting protein modules, or so-called reader domains, which can bind in a combinatorial modification-dependent manner. The enriched chromatin can be quantified by immunoblotting or mass spectrometry for the co-existence of histone modifications, while the associated DNA content can be analyzed by qPCR or next-generation sequencing. Altogether, MARCC provides a reproducible, efficient and customizable solution to enrich and analyze combinatorial histone modifications.
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