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Nucleosomes Exhibit Non‐uniform Unwrapping Along Native Chromatin Fibers with Increasing Salt Concentration as Revealed by Direct Imaging in a Microfluidic Channel
Author(s) -
Mori Hiroki,
Okeyo Kennedy O.,
Washizu Masao,
Oana Hidehiro
Publication year - 2018
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201700245
Subject(s) - chromatin , nucleosome , microfluidics , biophysics , chromosome , microbiology and biotechnology , fiber , dna , chemistry , nanotechnology , biology , materials science , genetics , gene , organic chemistry
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, the authors 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, the authors 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 are 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.

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