A-to-I RNA modifications in mouse oocytes

项目摘要/摘要 这项提议的主要目标是促进Brachova博士发展成为一名独立的 研究科学家。这一目标将通过积极的指导、教育相结合的方式来实现 培训、充实活动和研究。她的导师们很好地描述了导师阶段 包括每周联系和正式课程工作。研究的核心是岗位的作用-- 转录调控机制称为RNA编辑在整体卵母细胞质量中的作用。卵母细胞发育中的缺陷 减数分裂成熟是导致人类出生缺陷、流产和不孕的重要原因。这个 RNA转录后修饰的作用正在成为RNA稳定性的重要调节因素 和卵母细胞质量，然而作用于双链RNA(ADAR1)的腺苷脱氨酶的作用 催化腺苷(A)脱氨为肌苷(I)仍未被探索。这项研究的目的是 目的是确定ADAR1 RNA编辑对年轻和成熟卵母细胞RNA稳定性和卵母细胞功能的影响 老老鼠。初步数据显示，ADARFL/FLZP3-Cre卵母细胞减少了胚胎发育 这表明ADAR1是一个新的促进女性生育的因素。我们还表明， 来自老年人的卵母细胞和卵子降低了RNA编辑效率，这表明RNA编辑 缺陷可能与女性年龄相关的生育率下降有关。中心假设是 ADAR1 RNA编辑是一种通过调节RNA来维持雌性生育能力的新机制 卵母细胞生长和早期胚胎发育的稳定性。这一假设将分两个阶段进行检验。 独立的具体目标。特定目标1将评估ADAR1A-to-I RNA编辑在调节 母体在卵母细胞和早期胚胎中的mRNA剂量。特定目标2将决定ADAR1A-to-I RNA如何 编辑会影响卵母细胞中的mRNA降解。这项拟议的研究将利用幼崽的卵母细胞和卵子。 以及母体衰老的小鼠，以探索A-to-I编辑在卵子发生和与年龄相关的生育力降低中的作用。 这一建议代表了一种研究卵母细胞能力的创新方法，并具有扩展的潜力。 我们对女性生育能力的理解。此外，这里提出的研究是不可或缺的，以加强 对Brachova博士进行科学培训，使她能够实现实现独立和 成为女性生殖领域的独立科学家。

PROJECT SUMMARY/ABSTRACT The primary objective of this proposal is to facilitate the development of Dr. Brachova into an independent research scientist. This objective will be accomplished through a combination of active mentorship, didactic training, enrichment activities, and research. The mentorship phase is well described by her mentors and includes weekly contact and formal course work. The core of the research focuses on the role of a post- transcriptional regulation mechanism known as RNA editing in overall oocyte quality. Defects in oocyte growth and meiotic maturation represent a significant cause of human birth defects, miscarriage, and infertility. The contribution of RNA post-transcriptional modifications is emerging as an important regulator of RNA stability and oocyte quality, however the role of adenosine deaminase acting on double stranded RNA (ADAR1), which catalyzes the deamination of an adenosine (A) into inosine (I) remains unexplored. The goal of this research is to determine the effect of ADAR1 RNA editing on RNA stability and oocyte competence in young and old mice. Preliminary data show that ADARFL/FLZP3-Cre oocytes have reduced embryonic developmental potential, suggesting that ADAR1 is a novel factor that contributes to female fertility. We also show that oocytes and eggs from aged individuals have reduced RNA editing efficiency, suggesting that RNA editing deficiencies could be involved in female age-associated decline of fertility. The central hypothesis is that ADAR1 RNA editing is a novel mechanism for maintaining female fertility through the regulation of RNA stability during oocyte growth and early embryonic development. The hypothesis will be tested in two independent specific aims. Specific Aim 1 will assess the role of ADAR1 A-to-I RNA editing in the regulation of maternal mRNA dosage in the oocyte and early embryo. Specific Aim 2 will determine how ADAR1 A-to-I RNA editing effects mRNA degradation in oocytes. The proposed research will utilize oocytes and eggs from young and maternally aged mice to explore the role of A-to-I editing in oogenesis and age-related reduction in fertility. This proposal represents an innovative approach to study oocyte competence, and has the potential to expand our understanding of female fertility. Furthermore, the studies proposed herein are integral to enhance the scientific training of Dr. Brachova and enable her to accomplish her goal of achieving independence and becoming an independent scientist in the field of female reproduction.