Analysis of the Molecular Machinery Regulating Gene Expression during Vertebrate Development

脊椎动物发育过程中调控基因表达的分子机制分析

• 批准号: 10081285
• 负责人: Jean-Denis Beaudoin
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-28 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10081285
• 关键词: Academia; Address; Aging; Alleles; Animals; Appointment; Autoimmunity; Award; Biochemical; Biological Assay; Biological Phenomena; Biological Process; Biology; CRISPR/Cas technology; Code; Computer Analysis; Data; Defect; Deposition; Development; Developmental Biology; Educational process of instructing; Educational workshop; Elements; Embryo; Embryonic Development; Environment; Faculty; Fertilization; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genome; Goals; Health; Human; Human Development; In Vitro; Individual; Infertility; Inflammation; Learning; Life; Literature; Machine Learning; Malignant Neoplasms; Maternal Messenger RNA; Mediating; Mentors; Messenger RNA; Methods; Modeling; Molecular; Molecular Analysis; Monitor; Oocytes; Organism; Play; Polyribosomes; Post-Transcriptional Regulation; Postdoctoral Fellow; Property; Proteins; Prothrombin; RNA; RNA Conformation; RNA Helicase; RNA Probes; Reader; Regulation; Regulator Genes; Regulatory Element; Reporter; Reporter Genes; Research; Role; Scientist; Shapes; Signal Transduction; Structure; System; Techniques; Testing; Therapeutic; Training; Translations; Universities; Virus; Work; Zebrafish; analog; career; egg; experience; experimental study; human disease; in vivo; innovation; insight; loss of function; mRNA Decay; mRNA Stability; mutant; nervous system disorder; next generation sequencing; novel; novel strategies; programs; skills; sperm cell; tenure track; tool; transcriptome

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.

孕产妇至合子转变是所有动物中保守信息的基本传递， 其特征是转录景观的深刻变化。转录后调节中介 通过嵌入母体mRNA中的调节元件，基因表达的这种急剧变化。 RNA 结构不利于RNA功能和调节元素活性。基本生物学的协调 过程依赖于特定的RNA结构，例如RNA G-四链体介导的癌症翻译。 因此，该建议将解决生物学中的两个核心问题：该代码的组成部分是什么 调节早期胚胎发生以及单个组件在 脊椎动物的发展。新方法将用于了解这种重要的过渡。首先，高通量 将进行实验以识别调节mRNA丰度和翻译的mRNA元素（AIM 1），并解决其RNA结构（AIM 2）。然后，生化和功能性方法的结合将 用于发现这些监管元素的读者（AIM 3）。最后，这些读者的突变体将是 产生及其在脊椎动物发育过程中研究的分子机制和作用（AIM 3）。自从 RNA结构与读者之间的相互作用是关键生物学现象的主要调节剂（例如 炎症中的步态系统和自身免疫性的Roquin），新型基因表达调节网络 该提案中的揭露可能会在人类和影响人类的发展和健康中保存。到 完成这项建议，博多汤博士将继续他作为遗传学的博士后研究员的培训 耶鲁大学的系，他将同时享受最先进的设施以及与他的互动 该领域的导师和其他科学领袖。有了这个K99奖，博多博士的目标是结束 来自MRNA调节的几位科学专家（导师和合作者）的指导 生物学，机器学习和CRIS/CAS9介导的功能屏幕。此外，Beaudoin博士计划 通过参加结构化课程和讲习班来扩大他以前的教学和指导经验。 这将使他能够学习创新和有效的方法来教授生物学和进步以成为一个很好的 圆形的科学家和导师。 Beaudoin博士的现有专业知识以及此的科学和培训计划 提案将使他能够实现自己的长期职业目标：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 与数十种人类疾病有关（例如不育，神经系统疾病，癌症和衰老）。所以， 它们在相关脊椎动物模型中的分子表征将提供宝贵的见解以发展新的 人类治疗方法。 Beaudoin博士完全致力于获得任期的约会 学术界的初级教师。预计Beaudoin博士将对该小组负责人竞争 量身定制的K99/R00奖项完成后，旨在提高他的科学和指导技能。