Elucidation of mechanisms of germ cell reprogramming and establishment of the procedure to control epigenome.

阐明生殖细胞重编程机制并建立控制表观基因组的程序。

• 批准号: 21780269
• 负责人: INOUE Kimiko
• 金额: $ 2.83万
• 依托单位: The Institute of Physical and Chemical Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21780269/
• 关键词: 核移植クローン; エピゲノム; 遺伝子発現; X染色体; H3K9me2; DNAマイクロアレイ; Xist; 体細胞核移植クローン; 初期化; LOCKs; 体細胞クローン胚; Xist遺伝子

To improve understanding of the low efficiency of somatic cell nuclear transfer (SCNT), we performed global transcription analysis of mouse SCNT blastocysts. Many of the downregulated genes (21/39) identified in SCNT embryos were located on the X chromosome. When the downregulation of X-linked genes in SCNT embryos was corrected by using Xist-deficient donor cells, the birth rates increased 8- to 9-fold. Thus, SCNT may be significantly improved by correcting epigenetic errors specific for SCNT.In the next series of experiments, to understand the cause of consistent downregulation in the region of A7.2 and F3 on X chromosome (XqA7.2/F3) in SCNT blastocysts, we analyzed repressive modification, dimethylation of histone H3 lysine 9 (H3K9me2), with chromatin immunoprecipitation-on-chip data from cumulus cells. The results showed that H3K9me2 was broadly enriched in XqA7.2/F3 and formed tissue-specific blocks called as large organized chromatin K9 modification (LOCKs). These data suggested that downregulation of XqA7.2/F3 was caused by failure of reprogramming of LOCKs after nuclear transfer.

为了提高对体细胞核移植（SCNT）低效率的理解，我们对小鼠SCNT囊胚进行了整体转录分析。在SCNT胚胎中鉴定的许多下调基因（21/39）位于X染色体上。当使用Xist缺陷供体细胞纠正SCNT胚胎中X连锁基因的下调时，出生率增加了8至9倍。因此，通过纠正SCNT特有的表观遗传错误，SCNT可能会得到显着改善。在接下来的一系列实验中，为了了解SCNT囊胚中X染色体A7.2和F3区域（XqA7.2/F3）持续下调的原因，我们分析了组蛋白H3赖氨酸9（H3 K9 me 2）的抑制性修饰、二甲基化，以及来自卵丘细胞的染色质免疫沉淀芯片数据。结果表明，H3 K9 me 2在XqA7.2/F3中广泛富集，并形成组织特异性块，称为大组织化染色质K9修饰（LOCKs）。这些数据表明，XqA7.2/F3的下调是由核移植后LOCK重编程失败引起的。

项目成果

Sex-reversed somatic cell cloning in the mouse, 6th Annual Conference of Asian Reproductive Biotechnology Society 2009

小鼠性别逆转体细胞克隆，2009年亚洲生殖生物技术学会第六届年会

• 发表时间: 2009
• 作者: Inoue K;Ogonuki N;Mekada K;Yoshiki A;Sado T;OguraA
• 通讯作者: OguraA

Sex-Reversed Somatic Cell Cloning in the Mouse

• DOI: 10.1262/jrd.09-099e
• 发表时间: 2009-10-01
• 期刊: JOURNAL OF REPRODUCTION AND DEVELOPMENT
• 影响因子: 1.8
• 作者: Inoue, Kimiko;Ogonuki, Narumi;Ogura, Atsuo
• 通讯作者: Ogura, Atsuo

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Impeding Xist Expression from the Active X Chromosome Improves Mouse Somatic Cell Nuclear Transfer

• DOI: 10.1126/science.1194174
• 发表时间: 2010-10-22
• 期刊: SCIENCE
• 影响因子: 56.9
• 作者: Inoue, Kimiko;Kohda, Takashi;Ogura, Atsuo
• 通讯作者: Ogura, Atsuo

核移植によるリプログラミング,細胞42

通过核移植重编程，细胞 42

• 发表时间: 2010
• 作者: 小倉淳郎;井上貴美子
• 通讯作者: 井上貴美子