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: 9769091
• 负责人: Jean-Denis Beaudoin
• 金额: $ 11.54万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2020-02-27
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9769091
• 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; Structural Genes; 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.

母系到合子的转变是所有动物中保守的信息的基本转移， 其特点是转录景观发生了深刻的变化。转录后调节介导 这种基因表达的剧烈变化是通过嵌入母体mRNA中的调控元件实现的。RNA 结构对RNA功能和调节元件活性有害。协调基本生物 这些过程依赖于特定的RNA结构，例如癌症中RNA G-四链体介导的翻译。 因此，这个提议将解决生物学中的两个中心问题：密码的组成部分是什么 调节早期胚胎发生，以及在胚胎发育过程中各个成分的作用和分子功能是什么？ 脊椎动物发育。新的方法将被用来理解这一重要的转变。一是高吞吐量 将进行实验以鉴定调节mRNA丰度和翻译的mRNA元件（Aim 1），并解决其RNA结构（目标2）。然后，生物化学和功能方法的结合将 用于发现这些调控元件的读者（目标3）。最后，这些读者的突变体将 研究了它们在脊椎动物发育过程中的分子机制和作用（目的3）。以来 RNA结构和阅读器之间的相互作用是关键生物现象（例如， 炎症中的GAIT系统和自身免疫中的roquin），新的基因表达调控网络 该提案中所揭示的可能会在人类中保存并影响人类的发展和健康。到 完成这一建议，Beaudoin博士将继续他的培训作为博士后研究员在遗传学 他将在耶鲁大学的教育系，在那里他将享受最先进的设施和与他的同事的互动。 导师和该领域的其他科学领导者。凭借这个K99奖，Beaudoin博士的目标是接近 来自多位科学专家（导师和合作者）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 通过参加结构化的课程和研讨会来扩展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，成为一个很好的- 全面的科学家和导师Beaudoin博士现有的专业知识和科学和培训计划， 该提案将使他能够实现他的长期职业目标：建立一个研究计划来了解 RNA结构和RNA解旋酶在脊椎动物发育中的作用。缺陷RNA解旋酶已经被 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）。因此，我们认为， 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 学术界的初级教员完全可以预期，Beaudoin博士将有竞争力地担任这样的组长 K99/R 00奖完成后的任命旨在提高他的科学和指导技能。