Assessing the Effect of Chromatin-Binding Enzymes on Condensed Chromatin with Optical Tweezers

The formation of biomolecular condensates in vitro and in vivo has become an increasingly important subject of studies. One particular area of interest is the phase separation of chromatin in the nucleus. However, the interplay of condensed chromatin and chromatin-binding enzymes has barely been stu...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 2881; p. 345
Main Authors Kamp, Dieter, Vizjak, Petra, Stigler, Johannes
Format Journal Article
LanguageEnglish
Published United States 2025
Subjects
Biomolecular Condensates - chemistry
Biomolecular Condensates - metabolism
Chromatin - chemistry
Chromatin - metabolism
Chromatin Assembly and Disassembly
Humans
Optical Tweezers
Protein Binding
Chromatin
Condensates
Optical tweezers
Liquid-liquid phase separation (LLPS)
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Summary:The formation of biomolecular condensates in vitro and in vivo has become an increasingly important subject of studies. One particular area of interest is the phase separation of chromatin in the nucleus. However, the interplay of condensed chromatin and chromatin-binding enzymes has barely been studied as of now. Here, we show an optical tweezer-based assay that uses controlled fusion of two condensates to probe the effect of enzymes, such as chromatin remodeling motor proteins, on the fluidity of condensates. The assay provides a powerful tool that enables the study of if and how chromatin-enzyme interactions alter biophysical properties in these dense chromatin condensates.
ISSN:1940-6029
DOI:10.1007/978-1-0716-4280-1_17