Visualizing the Dynamics of Inactive X Chromosomes in Living Cells Using Antibody-Based Fluorescent Probes
Visualizing the Dynamics of Inactive X Chromosomes in Living Cells Using Antibody-Based Fluorescent Probes
复制标题
使用基于抗体的荧光探针可视化活细胞中非活性 X 染色体的动态
DOI：
10.1007/978-1-4939-8766-5_8
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发表时间：
2018
影响因子：
--
通讯作者：
Kimura Hiroshi
中科院分区：
文献类型：
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作者：
Sato Yuko;Stasevich Timothy J.;Kimura Hiroshi
失活X染色体（Xi）具有典型的表观遗传特征，包括组蛋白H3赖氨酸27三甲基化（H3K27me3）和H4赖氨酸20单甲基化（H4K20me1）的富集以及组蛋白乙酰化的缺乏。最近，这些修饰不仅在固定的标本中，而且通过来自修饰特异性抗体的探针在活细胞中被可视化。所述探针包括荧光标记的抗原结合片段（Fab），其可以被加载到细胞中，以及用绿色荧光蛋白标记的遗传编码的单链可变片段。我们将后者称为修饰特异性细胞内抗体，或简称“mintbodies”。通过使用Fab或mintbody靶向Xi特异性修饰，可以可视化活细胞中Xi的动力学。
The inactive X chromosome (Xi) harbors characteristic epigenetic features, including the enrichment of histone H3 lysine 27 trimethylation (H3K27me3) and H4 lysine 20 monomethylation (H4K20me1) as well as a lack of histone acetylation. Recently, these modifications have been visualized not only in fixed specimen, but also in living cells via probes derived from modification-specific antibodies. The probes include fluorescently labeled antigen binding fragments (Fabs), which can be loaded into cells, as well as genetically encoded single-chain variable fragments tagged with the green fluorescent protein. We refer to the latter as modification specific intracellular antibodies, or “mintbodies” for short. By using Fabs or mintbodies to target Xi-specific modifications, the dynamics of Xi in living cells can be visualized.