A-to-I RNA modifications in mouse oocytes

项目总结/摘要 该提案的主要目的是促进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、教育和培训相结合来实现。 培训、丰富活动和研究。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。研究的核心集中在一个职位的作用， 转录调控机制称为RNA编辑在整个卵母细胞质量。卵母细胞生长缺陷 和减数分裂成熟是人类出生缺陷、流产和不育的重要原因。的 RNA转录后修饰的贡献正在成为RNA稳定性的重要调节因子 然而，腺苷脱氨酶作用于双链RNA（ADAR 1）的作用， 催化腺苷（A）脱氨为肌苷（I）的方法尚未探索。本研究的目的 是为了确定ADAR 1 RNA编辑对年轻人RNA稳定性和卵母细胞能力的影响， 老老鼠初步数据显示，ADARFL/FLZP 3-Cre卵母细胞具有降低的胚胎发育 这表明ADAR 1是一种有助于女性生育的新因子。我们也证明了 来自老年个体的卵母细胞和卵子的RNA编辑效率降低，这表明RNA编辑 这些缺陷可能与女性年龄相关的生育能力下降有关。核心假设是， ADAR 1 RNA编辑是通过调节RNA维持女性生育力的新机制 卵母细胞生长和早期胚胎发育过程中的稳定性。假设将在两个 独立的具体目标。具体目标1将评估ADAR 1 A-to-I RNA编辑在调节细胞凋亡中的作用。 卵母细胞和早期胚胎中母体mRNA的剂量。具体目标2将确定ADAR 1 A到I RNA 编辑影响卵母细胞中的mRNA降解。这项研究将利用年轻人的卵母细胞和卵子 和母龄小鼠，以探索A-to-I编辑在卵子发生和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，并有可能扩大 我们对女性生育能力的理解此外，本文提出的研究是不可或缺的，以提高 Brachova博士的科学培训，使她能够实现她的目标，实现独立， 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.