Analysis of the Molecular Machinery Regulating Gene Expression during Vertebrate Development

母体向合子的转变是所有动物之间保存的信息的基本转移， 以转录格局的深刻变化为特征的。转录后调控介导 这种基因表达的急剧变化是通过嵌入母体mRNAs中的调控元件实现的。核糖核酸 结构不利于RNA的功能和调节元件的活性。基本生物学的协调 过程依赖于特定的RNA结构，如癌症中RNAG-四链介导的翻译。 因此，这项建议将解决生物学中的两个核心问题：密码的组成部分是什么 调控早期胚胎发育及其各个成分在胚胎发育中的作用和分子功能 脊椎动物的发育。将使用新的方法来理解这一至关重要的过渡。第一，高吞吐量 将进行实验以确定调节信使核糖核酸丰度和翻译的信使核糖核酸元件 1)，并解决它们的RNA结构(目标2)。然后，生物化学和功能方法的结合将 用于发现这些监管要素的读者(目标3)。最后，这些读者的变种将是 并研究了它们在脊椎动物发育过程中的分子机制和作用(目标3)。自.以来 RNA结构和阅读器之间的相互作用是关键生物现象的主要调节因素。 炎症中的步态系统和自身免疫中的roquin)，新的基因表达调控网络 在这项提案中发现的可能会被保存在人类身上，并影响人类的发展和健康。至 完成这项提议后，Beaudoin博士将继续他作为遗传学博士后研究员的培训 在耶鲁大学，他将享受最先进的设施和与他的 该领域的导师和其他科学领袖。有了这个K99奖，Beaudoin博士的目标是接近 来自几位科学专家(导师和合作者)的指导，在mRNA调控，发育 生物学、机器学习和CRISPR/Cas9介导的功能筛选。此外，Beaudoin博士计划 通过参加有组织的课程和研讨会，扩大他以前的教学和指导经验。 这将使他学会创新和有效地教授生物，并进步成为一名优秀的 全能的科学家和导师。Beaudoin博士现有的专业知识以及科学和培训计划 提案将使他能够实现他的长期职业目标：建立一个研究计划，以了解 RNA结构和RNA解旋酶在脊椎动物发育中的作用。有缺陷的RNA解旋酶已经被 与数十种人类疾病(如不孕不育、神经疾病、癌症和衰老)有关。因此， 它们在相关脊椎动物模型中的分子特征将为开发新的 人类的治疗方法。Beaudoin博士完全致力于获得终身教职 学术界的初级教员。人们完全预计博阿多因博士将成为这样的组长的竞争者 在他的K99/R00奖结束时的任命，以增强他的科学和指导技能。

The maternal to zygotic transition is a fundamental transfer of information conserved among all animals and characterized by a profound change of the transcriptional landscape. Post-transcriptional regulation mediates this drastic change in gene expression through regulatory elements embedded in maternal mRNAs. RNA structure is detrimental to RNA function and regulatory element activity. Coordination of essential biological processes relies on specific RNA structures such as RNA G-quadruplexes-mediated translation in cancer. Therefore, this proposal will address two central questions in biology: what are the components of the code regulating early embryogenesis and what is the role and molecular function of individual components in vertebrate development. New approaches will be used to understand this vital transition. First, high throughput experiments will be performed to identify mRNA elements that regulate mRNA abundance and translation (Aim 1), and to solve their RNA structure (Aim 2). Then, a combination of biochemical and functional approaches will be used to discover the readers of those regulatory elements (Aim 3). Finally, mutants of those readers will be generated and their molecular mechanism and role studied during vertebrate development (Aim 3). Since the interaction between RNA structures and readers are master regulators of key biological phenomenon (e.g. GAIT system in inflammation and roquin in autoimmunity), the novel gene expression regulatory networks uncover in this proposal will likely be conserved in human and impact human development and health. To accomplish this proposal, Dr. Beaudoin will continue his training as Postdoctoral fellow in the Genetics Department at Yale University, where he will enjoy both state-of-the-art facilities and the interaction with his mentors and other scientific leaders in the field. With this K99 Award, Dr. Beaudoin's goals are to get close mentoring from several scientific experts (mentors and collaborators) in mRNA regulation, developmental biology, machine learning and CRISPR/Cas9-mediated functional screens. Furthermore, Dr. Beaudoin plans to expand his previous teaching and mentoring experience by participating in structured courses and workshops. This will allow him to learn innovative and effective ways to teach biology and progress to become a well- rounded scientist and mentor. Dr. Beaudoin existing expertise and the scientific and training plans of this proposal will allow him to reaching his long-term career goal: to establish a research program to understand the role of RNA structures and RNA helicases in vertebrate development. Defective RNA helicases have been associated to dozens of human diseases (e.g. infertility, neurological disorders, cancers and aging). Therefore, their molecular characterization in a relevant vertebrate model will provide invaluable insights to develop new human therapeutic approaches. Dr. Beaudoin is fully committed to obtain an appointment as a tenure track junior faculty in academia. It is fully expected that Dr. Beaudoin will be competitive for such group leader appointments at the completion of his K99/R00 Award tailored to enhance his scientific and mentoring skills.