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纳米狭缝中单条天然染色质纤维的电泳拉伸与成像

Electrophoretic stretching and imaging of single native chromatin fibers in nanoslits.

作者信息

Yeh Jia-Wei, Szeto Kylan

机构信息

School of Applied and Engineering Physics, Cornell University, Ithaca, New York 14853, USA.

出版信息

Biomicrofluidics. 2017 Jul 25;11(4):044108. doi: 10.1063/1.4996340. eCollection 2017 Jul.

Abstract

Stretching single chromosomal DNA fibers in nanofluidic devices has become a valuable tool for studying the genome and more recently the epigenome. Although nanofluidic technology has been extensively used in single molecular DNA analysis, compared to bare DNA, much less work has been done to elongate chromatin, and only a few studies utilize more biologically relevant samples such as native eukaryotic chromatin. Here, we provide a method for stretching and imaging individual chromatin fibers within a micro- and nanofluidic device. This device was used to electrophoretically stretch and image single native chromatin fibers extracted from human cancer cells (HeLa cells) by attaching the chromatin to microspheres held at the entrance of a nanoslit. To further demonstrate the potential of this device in epigenetics, histone modification H3k79me2 was optically detected by fluorescence microscopy.

摘要

在纳米流体装置中拉伸单个染色体DNA纤维已成为研究基因组以及最近研究表观基因组的一种有价值的工具。尽管纳米流体技术已广泛应用于单分子DNA分析,但与裸DNA相比,在拉长染色质方面所做的工作要少得多,并且只有少数研究使用了更具生物学相关性的样本,如天然真核染色质。在此,我们提供了一种在微流控和纳米流控装置中拉伸和成像单个染色质纤维的方法。该装置用于通过将染色质附着到纳米狭缝入口处的微球上,对从人类癌细胞(HeLa细胞)中提取的单个天然染色质纤维进行电泳拉伸和成像。为了进一步证明该装置在表观遗传学中的潜力,通过荧光显微镜对组蛋白修饰H3k79me2进行了光学检测。

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本文引用的文献

1
Stretching of Tethered DNA in Nanoslits.纳米狭缝中束缚DNA的拉伸
ACS Macro Lett. 2016 Oct 18;5(10):1114-1118. doi: 10.1021/acsmacrolett.6b00639. Epub 2016 Sep 19.
3
Visualizing the entire DNA from a chromosome in a single frame.在单个框架中可视化染色体的整条 DNA。
Biomicrofluidics. 2015 Aug 5;9(4):044114. doi: 10.1063/1.4923262. eCollection 2015 Jul.
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Evolution. An epigenetic window into the past?进化。一扇通向过去的表观遗传之窗?
Science. 2014 Aug 1;345(6196):511-2. doi: 10.1126/science.1256515.
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Toward single-molecule optical mapping of the epigenome.朝向单分子光学表观基因组图谱绘制。
ACS Nano. 2014 Jan 28;8(1):14-26. doi: 10.1021/nn4050694. Epub 2013 Dec 20.

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