Biotechnology Journal
Biotechnology Journal
5 years ago

Measurement of nucleosomal structural stability distribution along individual giant native chromatin fibers at different salt concentrations using a microfluidic channel with microstructures

Kennedy O. Okeyo, Masao Washizu, Hiroki Mori, Hidehiro Oana
Identifying the distribution of the higher-order structure of chromatin—a complex of DNA and proteins—along genomic DNA can clarify the mechanisms underlying cell development and differentiation, including gene regulation. However, genome-wide analysis of this distribution at the single-cell level remains an outstanding challenge. Here, we report a new method for investigating changes in and the distribution of higher-order structures along native chromatin fibers—ranging over 100 μm in length—relative to changes in salt concentration. To this end, we developed a microfluidic platform that enabled us to isolate chromatin fibers from single cells and tether them to microstructures in a microfluidic channel without fragmentation. The fibers were then exposed to varying concentrations of salt solution under microscopic observation. As a result, the fibers were non-uniformly elongated by up to 2-3 times along the fiber axis as salt concentration was increased from 0 to 3 M, suggesting that chromosome structural stability is non-uniformly distributed along chromatin fibers in their native form. Thus, our system enables direct microscopic analysis of individual chromatin fibers from single cells, which can provide insights into epigenetic mechanisms of cell development, cell differentiation, and carcinogenesis.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/biot.201700245

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