Regulation of Polycomb by long noncoding RNAs during pre-implantation development

摘要 Polycomb Repressive Complex 2（PRC 2）是一种多亚基酶，在着床前发育中起重要作用。 每个主要胚胎谱系的发育和随后的发育。PRC 2抑制基因 表达，但其募集到特定基因组区域进行基因阻遏的机制仍然很差 明白已知PRC 2被抑制性长非编码RNA募集到特定的基因组区域 但目前还不清楚lncRNAs如何招募PRC 2并使其抑制基因， 有效地我将RNA结合蛋白（RBP）支架附着因子B1（SAFB 1）鉴定为PRC 2。 是PRC 2介导的基因沉默的重要因子。此外，先前的研究表明，SAFB 1 是发育所必需的，并在细胞应激期间起重要作用。我的提议将决定 SAFB 1、PRC 2和Xist之间相互作用的物理机制，RNA是否是SAFB 1/PRC 2的基础 特定的相互作用，最后，SAFB 1调节PRC 2的程度和这种相互作用所起的作用 在胚胎植入前发育的压力下。拟议项目的目标是确定PRC 2 在植入前发育中受抑制性lncRNA的调节，如果成功，将确定一种新的途径， 调节发育中胚胎的压力，并将提供关于PRC 2如何招募和调节的见解 通过lncRNA。随着时间的推移，这些实验可以为治疗性控制PRC 2的方法提供基础， 作为纠正结构性出生缺陷和改进体外受精方法的一种手段。 我的长期目标是成为一个独立的科学家在一个学术机构，我可以进行研究 在表观遗传学和发育的交叉点上。位于查佩尔山的北卡罗来纳州大学和 我的申办者实验室所在的药理学系是一个非常合作和支持的机构， 环境接受培训的技能，我需要达到我的目标。首先，在奖学金发放期间，我 计划接受广泛的实验技术培训，如基因组学，计算生物学，干细胞 生物学和发育表观遗传学，通过我的赞助商和共同赞助商的指导，以及通过 课程和冷泉港实验室的老鼠课程。第二，我计划发展一些技能， 获得科学独立。我将参加会议，并学习批判性地审查文献。第三，我要学会 通过发表至少一个第一作者的论文，清晰有效地传达我的科学，包括书面和口头的 在我的研究期间结束时发表和评论，并在各种公共场合展示我的研究成果。 论坛。最后，我将通过参加量身定制的职业研讨会来获得指导和领导技能。 为学术研究事业和科学教育。因此，我在这个奖项下的培训将导致 发展我实现长期目标所需的关键技能。

ABSTRACT Polycomb Repressive Complex 2 (PRC2) is a multi-subunit enzyme that is essential for preimplantation development and the subsequent development of every major embryonic lineage. PRC2 represses gene expression, yet the mechanisms of its recruitment to specific genomic regions for gene repression remain poorly understood. PRC2 is known to be recruited to specific genomic regions by repressive long non-coding RNAs (lncRNAs), such as Xist, but it is unclear how lncRNAs like recruit PRC2 and cause it to repress genes so effectively. I identified the RNA-binding protein (RBP) scaffold attachment factor B1 (SAFB1) as a PRC2 interactor and is important for PRC2 mediated gene silencing. Moreover, prior studies have shown that SAFB1 is essential for development and plays an important role during cellular stress. My proposal will determine the physical mechanism of interaction between SAFB1, PRC2, and Xist, whether RNA is the basis of SAFB1/PRC2 specific interactions, and finally, the extent to which SAFB1 regulates PRC2 and the roles this interaction plays during stress in preimplantation development. The objective of the proposed project is to determine how PRC2 is regulated by repressive lncRNAs in preimplantation development and if successful, will identify a new pathway that regulates stress in the developing embryo and will provide insight as to how PRC2 is recruited and regulated by lncRNAs. In time, these experiments can provide a foundation for approaches to control PRC2 therapeutically, as a means to correct structural birth defects, and to improve methods for in vitro fertilization. My long-term goal is to become an independent scientist at an academic institution where I can perform research at the intersection of epigenetics and development. The University of North Carolina at Chapel Hill, and the Department of Pharmacology in which my sponsor’s lab is housed, is an extremely collaborative and supportive environment to receive training in the skills I need to attain my goal. First, during the fellowship award period, I plan receive extensive training in experimental techniques such as genomics, computational biology, stem cell biology, and developmental epigenetics, through guidance from my sponsor and co-sponsor, as well as through classes and a Cold Spring Harbor Laboratories mouse course. Second, I plan to develop skills that will allow me to gain scientific independence. I will attend meetings and learn to critically review literature. Third, I will learn to clearly and effectively communicate my science, written and orally, through publishing at least one first authored publication and review by the end of my fellowship period, and by presenting my research at various public forums. Lastly, I will gain skills in mentorship and leadership through participation in career workshops tailored for academic research careers and science education. Therefore, my training under this award will result in the development of crucial skills needed for me to achieve my long-term goal.