Genetically encoded system to track histone modification in vivo.
Genetically encoded system to track histone modification in vivo.
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DOI：
10.1038/srep02436
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发表时间：
2013
影响因子：
4.6
通讯作者：
Kimura, Hiroshi
中科院分区：
文献类型：
--
作者：
Sato, Yuko;Mukai, Masanori;Ueda, Jun;Muraki, Michiko;Stasevich, Timothy J.;Horikoshi, Naoki;Kujirai, Tomoya;Kita, Hiroaki;Kimura, Taisuke;Hira, Seiji;Okada, Yasushi;Hayashi-Takanaka, Yoko;Obuse, Chikashi;Kurumizaka, Hitoshi;Kawahara, Atsuo;Yamagata, Kazuo;Nozaki, Naohito;Kimura, Hiroshi
翻译后的组蛋白修饰在基因调节、发育和分化中起着关键作用，但它们在生物体中的动态变化几乎完全未知。为了解决这个问题，我们开发了一种基因编码系统，通过产生可在体内表达的荧光修饰特异性细胞内抗体(微抗体)来跟踪组蛋白的修饰。为了证明这一点，我们设计了一种H3赖氨酸9乙酰化特异性薄荷抗体(H3K9ac-薄荷抗体)，并在人细胞中稳定表达。与其目标乙酰化的定位很好地一致，H3K9ac-薄荷体富含常染色质，其动力学在组蛋白脱乙酰酶抑制剂处理后发生了可测量的变化。我们还获得了稳定表达H3K9ac-薄荷抗体的转基因果蝇和斑马鱼，用于体内示踪。在果蝇胚胎发育过程中，H3K9抗体定位的戏剧性变化可能突出了在有丝分裂周期7附近合子转录开始时乙酰化的增强。总之，这项工作证明了薄荷抗体的广泛潜力，并为体内的表观遗传学分析奠定了基础。
Post-translational histone modifications play key roles in gene regulation, development, and differentiation, but their dynamics in living organisms remain almost completely unknown. To address this problem, we developed a genetically encoded system for tracking histone modifications by generating fluorescent modification-specific intracellular antibodies (mintbodies) that can be expressed in vivo. To demonstrate, an H3 lysine 9 acetylation specific mintbody (H3K9ac-mintbody) was engineered and stably expressed in human cells. In good agreement with the localization of its target acetylation, H3K9ac-mintbody was enriched in euchromatin, and its kinetics measurably changed upon treatment with a histone deacetylase inhibitor. We also generated transgenic fruit fly and zebrafish stably expressing H3K9ac-mintbody for in vivo tracking. Dramatic changes in H3K9ac-mintbody localization during Drosophila embryogenesis could highlight enhanced acetylation at the start of zygotic transcription around mitotic cycle 7. Together, this work demonstrates the broad potential of mintbody and lays the foundation for epigenetic analysis in vivo.
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影响因子：
2.4
作者：
Kvam, Erik;Sierks, Michael R.;Messer, Anne
通讯作者：
Messer, Anne
影响因子：
48
作者：
Rothbauer, Ulrich;Zolghadr, Kourosh;Leonhardt, Heinrich
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Leonhardt, Heinrich
影响因子：
4.6
作者：
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通讯作者：
Lipshitz, Howard D.
影响因子：
14.9
作者：
ZHOU, H;FISHER, RJ;PAPAS, TS
通讯作者：
PAPAS, TS
影响因子：
4.8
作者：
Asakawa, Kazuhide;Kawakami, Koichi
通讯作者：
Kawakami, Koichi