Tracking epigenetic histone modifications in single cells using Fab-based live endogenous modification labeling

Yoko Hayashi-Takanaka, Kazuo Yamagata, Teruhiko Wakayama, Timothy J. Stasevich, Takashi Kainuma, Toshiki Tsurimoto, Makoto Tachibana, Yoichi Shinkai, Hitoshi Kurumizaka, Naohito Nozaki, Hiroshi Kimura

研究成果: ジャーナルへの寄稿記事

120 引用 (Scopus)

抄録

Histone modifications play an important role in epigenetic gene regulation and genome integrity. It remains largely unknown, however, how these modifications dynamically change in individual cells. By using fluorescently labeled specific antigen binding fragments (Fabs), we have developed a general method to monitor the distribution and global level of endogenous histone H3 lysine modifications in living cells without disturbing cell growth and embryo development. Fabs produce distinct nuclear patterns that are characteristic of their target modifications. H3K27 trimethylation-specific Fabs, for example, are concentrated on inactive X chromosomes. As Fabs bind their targets transiently, the ratio of bound and free molecules depends on the target concentration, allowing us to measure changes in global modification levels. High-affinity Fabs are suitable for mouse embryo imaging, so we have used them to monitor H3K9 and H3K27 acetylation levels in mouse preimplantation embryos produced by in vitro fertilization and somatic cell nuclear transfer. The data suggest that a high level of H3K27 acetylation is important for normal embryo development. As Fab-based live endogenous modification labeling (FabLEM) is broadly useful for visualizing any modification, it should be a powerful tool for studying cell signaling and diagnosis in the future.

元の言語英語
ページ(範囲)6475-6488
ページ数14
ジャーナルNucleic acids research
39
発行部数15
DOI
出版物ステータス出版済み - 8 1 2011

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Histone Code
Epigenomics
Acetylation
Embryonic Development
X Chromosome
Blastocyst
Fertilization in Vitro
Growth and Development
Histones
Lysine
Embryonic Structures
Genome
Antigens
Genes

All Science Journal Classification (ASJC) codes

  • Genetics

これを引用

Hayashi-Takanaka, Y., Yamagata, K., Wakayama, T., Stasevich, T. J., Kainuma, T., Tsurimoto, T., ... Kimura, H. (2011). Tracking epigenetic histone modifications in single cells using Fab-based live endogenous modification labeling. Nucleic acids research, 39(15), 6475-6488. https://doi.org/10.1093/nar/gkr343

Tracking epigenetic histone modifications in single cells using Fab-based live endogenous modification labeling. / Hayashi-Takanaka, Yoko; Yamagata, Kazuo; Wakayama, Teruhiko; Stasevich, Timothy J.; Kainuma, Takashi; Tsurimoto, Toshiki; Tachibana, Makoto; Shinkai, Yoichi; Kurumizaka, Hitoshi; Nozaki, Naohito; Kimura, Hiroshi.

:: Nucleic acids research, 巻 39, 番号 15, 01.08.2011, p. 6475-6488.

研究成果: ジャーナルへの寄稿記事

Hayashi-Takanaka, Y, Yamagata, K, Wakayama, T, Stasevich, TJ, Kainuma, T, Tsurimoto, T, Tachibana, M, Shinkai, Y, Kurumizaka, H, Nozaki, N & Kimura, H 2011, 'Tracking epigenetic histone modifications in single cells using Fab-based live endogenous modification labeling', Nucleic acids research, 巻. 39, 番号 15, pp. 6475-6488. https://doi.org/10.1093/nar/gkr343
Hayashi-Takanaka, Yoko ; Yamagata, Kazuo ; Wakayama, Teruhiko ; Stasevich, Timothy J. ; Kainuma, Takashi ; Tsurimoto, Toshiki ; Tachibana, Makoto ; Shinkai, Yoichi ; Kurumizaka, Hitoshi ; Nozaki, Naohito ; Kimura, Hiroshi. / Tracking epigenetic histone modifications in single cells using Fab-based live endogenous modification labeling. :: Nucleic acids research. 2011 ; 巻 39, 番号 15. pp. 6475-6488.
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