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

摘要 多梳抑制复合体2(Prc2)是一种多亚单位酶，在着床前是必不可少的。 每一个主要胚胎谱系的发育和随后的发育。PrC2抑制基因 表达，但其募集到特定基因组区域以进行基因抑制的机制仍然很差 明白了。已知PRC2通过抑制性的长非编码RNA被招募到特定的基因组区域 (LncRNAs)，如Xist，但目前尚不清楚像这样的lncRNAs是如何招募PRC2并导致其抑制基因的 有效地。我鉴定了RNA结合蛋白(RBP)支架附着因子B1(SAFB1)是一个PRC2 在PRC2介导的基因沉默中起重要作用。此外，先前的研究表明，SAFB1 是发育所必需的，在细胞应激过程中起着重要作用。我的提案将决定 SAFB1、PRC2和XIST相互作用的物理机制，RNA是否是SAFB1/PRC2的基础 具体的相互作用，最后，SAFB1对PRC2的监管程度和这种相互作用所起的作用 在植入前发育的应激过程中。拟议项目的目标是确定PRC2如何 在植入前发育中受抑制性lncRNA调控，如果成功，将发现一条新的途径 它调节发育中的胚胎的压力，并将提供关于PRC2是如何被招募和调节的洞察力 通过InncRNAs。随着时间的推移，这些实验可以为从治疗上控制PRC2的方法提供基础， 作为纠正结构性出生缺陷的手段，并改进体外受精的方法。 我的长期目标是成为一名学术机构的独立科学家，在那里我可以进行研究 处于表观遗传学和发育的交汇点。北卡罗来纳大学教堂山分校和 我的赞助商的实验室所在的药理学系，是一个非常合作和支持的 我需要一个环境来接受技能培训，以达到我的目标。首先，在奖学金获得期内，我 Plans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。其次，我计划发展一些技能，使我能够 以获得科学上的独立。我将参加会议，并学会批判性地复习文献。第三，我要学会 通过发表至少一篇第一作者的文章，清楚而有效地传播我的书面和口头科学 在我的研究期结束时发表和评论，并通过在不同的公众面前展示我的研究 论坛。最后，我将通过参加量身定做的职业研讨会来获得指导和领导技能 用于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 培养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.