追踪染色质修饰动力学的活细胞成像探针

IF 1.8 4区 工程技术
Microscopy Pub Date : 2021-10-01 DOI:10.1093/jmicro/dfab030
Yuko Sato;Masaru Nakao;Hiroshi Kimura
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引用次数: 8

摘要

染色质的时空组织在细胞核中受到不同水平的调控。表观遗传学修饰,如DNA甲基化和组蛋白修饰,参与染色质调节,并在基因组功能中发挥基本作用。虽然染色质免疫沉淀和测序已经揭示了许多细胞类型中的一维表观基因组景观,但染色质修饰的动态变化及其与染色质组织和基因组功能的相关性仍然难以捉摸。观察染色质及其修饰的活细胞探针已成为监测染色质动态调节的强大工具。体积染色质可以通过小荧光染料和荧光蛋白进行可视化,并且通过调整基因组编辑工具已经检测到特定的内源性基因组基因座。为了追踪活细胞中的染色质修饰,已经开发了各种类型的探针。与特定修饰结合较弱的蛋白质结构域,如组蛋白甲基化的色域,可以重复产生更紧密的结合探针,然后用荧光蛋白标记。还证明了来自修饰特异性抗体的抗原结合片段和单链可变片段可以在不干扰细胞分裂、发育和分化的情况下用作结合探针。这些修饰结合模块用于基于荧光/Förster共振能量转移的修饰传感器,以测量由修饰引发的分子内构象变化。其他探针可以使用二价结合系统产生,例如荧光互补或萤光素酶化学发光。使用这些探针的活细胞染色质修饰成像将解决染色质的动态调节问题,并将有助于分析和筛选细胞和生物体中有效的表观基因组药物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Live-cell imaging probes to track chromatin modification dynamics

Live-cell imaging probes to track chromatin modification dynamics

Live-cell imaging probes to track chromatin modification dynamics

Live-cell imaging probes to track chromatin modification dynamics
The spatiotemporal organization of chromatin is regulated at different levels in the nucleus. Epigenetic modifications such as DNA methylation and histone modifications are involved in chromatin regulation and play fundamental roles in genome function. While the one-dimensional epigenomic landscape in many cell types has been revealed by chromatin immunoprecipitation and sequencing, the dynamic changes of chromatin modifications and their relevance to chromatin organization and genome function remain elusive. Live-cell probes to visualize chromatin and its modifications have become powerful tools to monitor dynamic chromatin regulation. Bulk chromatin can be visualized by both small fluorescent dyes and fluorescent proteins, and specific endogenous genomic loci have been detected by adapting genome-editing tools. To track chromatin modifications in living cells, various types of probes have been developed. Protein domains that bind weakly to specific modifications, such as chromodomains for histone methylation, can be repeated to create a tighter binding probe that can then be tagged with a fluorescent protein. It has also been demonstrated that antigen-binding fragments and single-chain variable fragments from modification-specific antibodies can serve as binding probes without disturbing cell division, development and differentiation. These modification-binding modules are used in modification sensors based on fluorescence/Förster resonance energy transfer to measure the intramolecular conformational changes triggered by modifications. Other probes can be created using a bivalent binding system, such as fluorescence complementation or luciferase chemiluminescence. Live-cell chromatin modification imaging using these probes will address dynamic chromatin regulation and will be useful for assaying and screening effective epigenome drugs in cells and organisms.
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来源期刊
Microscopy
Microscopy 工程技术-显微镜技术
自引率
11.10%
发文量
0
审稿时长
>12 weeks
期刊介绍: Microscopy, previously Journal of Electron Microscopy, promotes research combined with any type of microscopy techniques, applied in life and material sciences. Microscopy is the official journal of the Japanese Society of Microscopy.
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