The role of chromatin modifications in the efficiency of reprogramming

染色质修饰在重编程效率中的作用

• 批准号: 66393331
• 负责人: Professor Dr. Konstantinos Anastassiadis
• 金额: --
• 依托单位: Biotechnologisches Zentrum (Biotec)
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2008
• 资助国家: 德国
• 起止时间: 2007-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/66393331?language=en
• 关键词: role; chromatin; modifications; efficiency; reprogramming

The molecular mechanisms involved in reprogramming of a differentiated mammalian cell to an embryonic like pluripotent cell are poorly defined. In the first funding period we established a transposon based induced reprogramming assay using Neural Stem (NS) cells and analysed the role of histone methyltransferases and histone demethylases. Conditional mutagenesis of the methyltransferase Mll1 in NS cells resulted in a 8-fold increase of induced pluripotent stem cell (iPS) colonies. In contrast a reduction in the number of iPS colonies was observed after conditional inactivation of Mll2. We also obtained more iPS colonies after tetracycline-regulated overexpression of the histone demethylases Jmjd1a (H3K9me2) and Utx (H3K27me3). In the second funding period we aim at identifying the targets of the above-mentioned histone methyltransferases and demethylases in NS cells as a first step for unravelling the molecular mechanisms of reprogramming. Furthermore we will continue our efforts for generating conditional mutagenesis and inducible expression/RNAi systems for the other existing demethylases and extend the analysis using in addition to induced reprogramming assays, ES-cell fusion and nuclear transfer assays.

将分化的哺乳动物细胞重新编程为胚胎样多能细胞所涉及的分子机制尚不清楚。在第一个资助期，我们建立了一种基于转座子的神经干细胞诱导重编程实验，并分析了组蛋白甲基转移酶和组蛋白去甲基酶的作用。在NS细胞中对甲基转移酶M111进行条件性突变后，诱导的多能干细胞(IPS)集落数增加了8倍。相反，在条件灭活MLL2后观察到iPS集落数量的减少。在四环素调控的组蛋白去甲基酶Jmjd1a(H3K9me2)和UTx(H3K27me3)过表达后，我们也获得了更多的iPS克隆。在第二个资助期，我们的目标是确定上述NS细胞中组蛋白甲基转移酶和去甲基酶的靶点，作为解开重编程分子机制的第一步。此外，我们将继续努力为其他现有的脱甲基酶建立条件突变和可诱导表达/RNAi系统，并扩大分析范围，除诱导重编程分析外，还使用ES细胞融合和核转移分析。

项目成果

The H3K4 methyltransferase Setd1a is first required at the epiblast stage, whereas Setd1b becomes essential after gastrulation

• DOI: 10.1242/dev.098152
• 发表时间: 2014-03-01
• 期刊: DEVELOPMENT
• 影响因子: 4.6
• 作者: Bledau, Anita S.;Schmidt, Kerstin;Anastassiadis, Konstantinos
• 通讯作者: Anastassiadis, Konstantinos

MLL2 Is Required in Oocytes for Bulk Histone 3 Lysine 4 Trimethylation and Transcriptional Silencing

• DOI: 10.1371/journal.pbio.1000453
• 发表时间: 2010-08-01
• 期刊: PLOS BIOLOGY
• 影响因子: 9.8
• 作者: Andreu-Vieyra, Claudia V.;Chen, Ruihong;Matzuk, Martin M.
• 通讯作者: Matzuk, Martin M.