P0044380 Parchem - Role of Small RNA's in Development

P0044380 Parchem - 小RNA在发育中的作用

• 批准号: 8471017
• 负责人: Ronald J Parchem
• 金额: $ 5.57万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-30 至 2014-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8471017
• 关键词: Biological; Biology; Bypass; Cells; Defect; Development; Embryo; Embryonic Development; Epiblast; Extraembryonic Structure; Future; Germ Layers; Human; Lead; Mediating; Methods; MicroRNAs; Molecular; Molecular Target; Mus; Pathway interactions; Pattern; Pluripotent Stem Cells; Process; Regenerative Medicine; Role; Scientist; Seeds; Somatic Cell; Techniques; Testing; Therapeutic; Therapeutic Uses; Tissues; Transcript; embryonic stem cell; functional restoration; gastrulation; human disease; in vivo; pluripotency; research study; stem cells

DESCRIPTION (provided by applicant): Embryonic stem cells and the molecular methods to induce their formation from somatic cells has been the subject of intense study in recent years. These techniques allow scientists to bypass the ethically contentious issue of using human embryos and create personalized pluripotent stem cells. These advances underlie the incredible push to begin using these cells for therapeutic purposes. Unfortunately, our ability to generate cells or tissues for therapeutic use is still quite limited. The Blelloch lab has recently shown tht miRNAs are necessary for differentiation of embryonic stem cells, and that specific classes of miRNAs function through opposing pathways to mediate differentiation out of the pluripotent state. This finding suggests that miRNAs are important global regulators of differentiation. In thi application, I hypothesize that the mir302 cluster of miRNAs is critical in the transition from pluripotency to differentiated germ layers by regulating transcripts that mechanistically underlie this critically important cell-fate decision. To test this hypothesis I will first determine the invivo role of mir302 miRNAs by characterizing epiblast formation and gastrulation during embryonic development of mir302 deficient mice. I will then characterize differentiation defects in mouse embryonic stem cells lacking mir302. Next, I will identify and functionally test the molecular targets of mir302 to identify the mechanism through which mir302 regulates germ layer specification. Finally, I will dissect the function of miRNAs containing different seed sequences within the mir302 locus to understand how co- expressed miRNAs with different targets function in development. These experiments will provide detailed information on how miRNAs regulate cell fate decisions and through what mechanism a cell transitions from pluripotency to the primary embryonic germ layers.

描述（由申请人提供）：近年来，胚胎干细胞和诱导其从体细胞形成的分子方法一直是近年来的强烈研究的主题。这些技术使科学家能够绕过使用人类胚胎并创建个性化多能干细胞的道德上有争议的问题。这些进步是令人难以置信的推动，以开始将这些细胞用于治疗目的。不幸的是，我们生成细胞或组织用于治疗的能力仍然非常有限。 Blelloch Lab最近表明，MiRNA对于分化胚胎干细胞是必需的，并且通过相反的途径介导从多能状态的介导分化的途径，特定类别的miRNA功能。这一发现表明，miRNA是分化的重要全球调节因子。在应用中，我假设miR302 miRNA簇在从多能性到分化的细菌层的过渡中至关重要，通过调节这些非常重要的细胞命中率决策的转录本机构基础。为了检验这一假设，我将首先通过表征MiR302缺乏小鼠的胚胎发育过程中表征层状形成和胃肠道来确定miR302 miRNA的无视作用。然后，我将表征缺乏miR302的小鼠胚胎干细胞中的分化缺陷。接下来，我将识别并在功能上测试miR302的分子靶标，以确定miR302调节细菌层规范的机制。最后，我将剖析miR302基因座中包含不同种子序列的miRNA功能，以了解如何在发育中与不同靶标发挥作用的miRNA。这些实验将提供有关miRNA如何调节细胞命运决策的详细信息，以及通过细胞从多能性到主要胚胎细菌层的机理转变。

项目成果

BAC library for the amphipod crustacean, Parhyale hawaiensis.

• DOI: 10.1016/j.ygeno.2010.03.005
• 发表时间: 2010-05
• 期刊: GENOMICS
• 影响因子: 4.4
• 作者: Parchem, Ronald J.;Poulin, Francis;Stuart, Andrew B.;Amemiya, Chris T.;Patel, Nipam H.
• 通讯作者: Patel, Nipam H.