Genomic-wide Analysis of Oct 3/4 and Nanog Targets

Oct 3/4 和 Nanog 目标的全基因组分析

基本信息

批准号：
7455921
负责人：
SOREN IMPEY
金额：
$ 28.41万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-07-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7455921
关键词：
Address Adult Animals Attention Binding Binding Sites Biological Assay Carcinoma Cataloging Catalogs Cell Line Cell Therapy Cell division Cells Chromatin Code Communities Complement Complex DNA Degenerative Disorder Deposition Development Diabetes Mellitus Embryo Embryonic Development Enhancers Epigenetic Process Excision Functional RNA Generations Genes Genome Genomics Growth Histones Inner Cell Mass Knowledge Lead Libraries Light Lysine Mediating Messenger RNA Methods Methylation Methyltransferase MicroRNAs Modification Molecular Mus Nature Parkinson Disease Pathway interactions Play Pluripotent Stem Cells Polycomb Polymerase Chain Reaction Principal Investigator Promoter Regions Proteins Recruitment Activity Regulation Regulon Repression Research Design Retinoids Role Site Small Interfering RNA Stem Cell Research Stem cells System Thinking Time Tissues Transcript Work base blastomere structure cell growth chromatin immunoprecipitation demethylation embryonic stem cell feeding high throughput analysis homeodomain human embryonic stem cell human stem cells imprint insight natural Blastocyst Implantation novel novel strategies pluripotency programs self-renewal stem stem cell therapy transcription factor

项目摘要

DESCRIPTION (provided by applicant): Embryonic stem (ES) cells isolated from the inner cell mass of murine blostocysts are pluripotent, capable of indefinite symmetric cell division, and can generate chimeric animals. The recent isolation of human ES cells holds great promise for the treatment of a variety of degenerative disorders including, but not limited to, Parkinson's disease and diabetes. The homeodomain transcription factors, Oct3/4 and Nanog, are believed to play critical roles in sustaining ES cell pluripotency. Surprisingly, targets of Nanog are completely unknown and few Oct3/4 targets have been proposed. Thus, the molecular mechanisms that repress differentiation of ES cells and promote self-renewal are poorly defined. We have developed a novel approach for identifying complex metazoan regulons called SACO (Serial Analysis of Chromatin Occupancy), which combines chromatin immunoprecipitation with a modification of Long SAGE. We will use SACO to identify the entire complement of Nanog and Oct3/4 genomic targets in mouse ES cells. Such knowledge would not only aid efforts to characterize mechanisms that govern cell-fate commitment of stem cells, but could also enable the indefinite propagation of existing human stem cell lines. Insight into these transcriptional networks could also lead to the generation of pluripotent stem cells from adult tissues or cells. Our studies are designed to provide a complete definition of Nanog and Oct3/4 targets in pluripotent stem cells. By characterizing the regulation of the corresponding transcripts, we expect to characterize the molecular pathways that control self-renewal and pluripotency of ES cells. The microarrays representing novel Nanog and Oct3/4 driven transcripts and the catalog of genomic binding sites will be provided to the stem cell research community. The creation of microarrays representing novel Nanog and Oct3/4 targets will facilitate high-throughput analysis of their expression during early embryogenesis and their regulation by pathways that regulate differentiation and pluripotency. This set of studies will represent the most comprehensive analysis to date of the regulation of transcription factor binding in any metazoan system.

描述（由申请人提供）：从小鼠囊胚内细胞团分离的胚胎干（ES）细胞是多能的，能够无限期对称细胞分裂，并且可以产生嵌合动物。最近分离的人类 ES 细胞为治疗各种退行性疾病（包括但不限于帕金森病和糖尿病）带来了巨大希望。同源结构域转录因子 Oct3/4 和 Nanog 被认为在维持 ES 细胞多能性方面发挥着关键作用。令人惊讶的是，Nanog 的目标完全未知，并且很少有人提出 Oct3/4 目标。因此，抑制 ES 细胞分化和促进自我更新的分子机制尚不清楚。我们开发了一种识别复杂后生动物调节子的新方法，称为 SACO（染色质占用序列分析），该方法将染色质免疫沉淀与 Long SAGE 的修饰相结合。我们将使用 SACO 来鉴定小鼠 ES 细胞中 Nanog 和 Oct3/4 基因组靶标的完整补充。这些知识不仅有助于描述控制干细胞细胞命运承诺的机制，而且还可以实现现有人类干细胞系的无限繁殖。对这些转录网络的深入了解还可能导致从成体组织或细胞产生多能干细胞。我们的研究旨在提供多能干细胞中 Nanog 和 Oct3/4 靶标的完整定义。通过表征相应转录本的调节，我们期望表征控制 ES 细胞自我更新和多能性的分子途径。代表新型 Nanog 和 Oct3/4 驱动转录本的微阵列以及基因组结合位点目录将提供给干细胞研究界。代表新的 Nanog 和 Oct3/4 靶标的微阵列的创建将有助于对它们在早期胚胎发生过程中的表达及其通过调节分化和多能性的途径的调节进行高通量分析。这组研究将代表迄今为止对任何后生动物系统中转录因子结合调节的最全面的分析。