Induction of pluripotency by protein gain of function in cloned mouse embryos

克隆小鼠胚胎中蛋白质功能获得诱导多能性

• 批准号: 66209674
• 负责人: Privatdozent Dr. Michele Boiani
• 金额: --
• 依托单位: Abteilung Zell- und Entwicklungsbiologie
• 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/66209674?language=en
• 关键词: Induction; pluripotency; protein; gain; function

Compared to the other reprogramming methods, somatic cell nuclear transfer into oocytes relies on a naturally existing cell, and is fast and totipotent. Since the resultant pluripotent stem cells are of superior quality, and many oocytic mRNAs find no match in the oocyte proteome, we put forward the hypotheses that: 1) the oocytic reprogramming factors differ from those known in ES and iPS cells; and 2) oocyte-mediated reprogramming uses also new proteins that are General information synthesized immediately after oocyte activation. We predict these oocytic factors to synergize with the known iPS reprogramming factors, leading to a more complete epigenetic 'resetting' of the somatic genome. Accordingly, we propose two sets of experiments. Firstly, we will combine biochemistry, mass spectrometry and bioinformatics to identify candidate reprogramming factors present in the pronuclei of activated mouse oocytes. Objective screening criteria have already been tested on the proteome of metaphase II oocytes, retaining 29 of 3699 proteins. Secondly, we will express the old (29) and new candidates in mouse fibroblasts during their conversion to iPS cells. The expected gain in reprogramming efficiency will be assessed by comparing rates of Nanog gene induction and pluripotent cell colony formation in the presence vs absence of the oocytic factor. Our project is very innovative on at least three fronts: firstly, it will define the catalog of natural reprogramming factors that operate in the oocyte. Secondly, our project will be the first oocyteeducated attempt to find the limiting stage of iPS reprogramming. Thirdly, independently of reprogramming function, our list of proteins will bring new insight in the molecular landscape of the mature female germ cell.

与其他重编程方法相比，体细胞核移植到卵母细胞中依赖于天然存在的细胞，并且是快速和全能的。由于所得到的多能干细胞具有上级质量，并且许多卵细胞mRNA在卵母细胞蛋白质组中找不到匹配，因此我们提出以下假设：1）卵母细胞重编程因子不同于ES和iPS细胞中已知的那些因子;以及2）卵母细胞介导的重编程也使用新的蛋白质，这些蛋白质是卵母细胞活化后立即合成的一般信息。我们预测这些卵细胞因子与已知的iPS重编程因子协同作用，导致体细胞基因组更完整的表观遗传“重置”。因此，我们提出了两组实验。首先，我们将结合联合收割机生物化学、质谱和生物信息学来鉴定激活的小鼠卵母细胞原核中存在的候选重编程因子。目的筛选标准已经在中期II卵母细胞的蛋白质组上进行了测试，保留了3699种蛋白质中的29种。其次，我们将在小鼠成纤维细胞转化为iPS细胞的过程中表达旧的（29）和新的候选物。将通过比较存在与不存在卵细胞因子的情况下Nanog基因诱导和多能细胞集落形成的速率来评估重编程效率的预期增益。我们的项目至少在三个方面非常创新：首先，它将定义在卵母细胞中运作的天然重编程因子的目录。其次，我们的项目将是第一个卵母细胞培养的尝试，以找到iPS重编程的限制阶段。第三，独立于重编程功能，我们的蛋白质列表将为成熟女性生殖细胞的分子景观带来新的见解。

项目成果

Maternal age effect on mouse oocytes: new biological insight from proteomic analysis.

• DOI: 10.1530/rep-14-0126
• 发表时间: 2014-07
• 期刊: Reproduction
• 影响因子: 3.8
• 作者: C. Schwarzer;Marcin Siatkowski;M. Pfeiffer;Nicole Baeumer;H. Drexler;Bingyuan Wang;G. Fuellen;M. Boia

Somatic cell nuclear reprogramming of mouse oocytes endures beyond reproductive decline

• DOI: 10.1111/j.1474-9726.2010.00644.x
• 发表时间: 2011-02-01
• 期刊: AGING CELL
• 影响因子: 7.8
• 作者: Esteves, Telma Cristina;Balbach, Sebastian Thomas;Boiani, Michele
• 通讯作者: Boiani, Michele

Differences in embryo quality are associated with differences in oocyte composition: A proteomic study in inbred mice

• DOI: 10.1002/pmic.201400334
• 发表时间: 2015-02-01
• 期刊: PROTEOMICS
• 影响因子: 3.4
• 作者: Pfeiffer, Martin J.;Taher, Leila;Boiani, Michele
• 通讯作者: Boiani, Michele