Regulating RNA function by modulating RNA folding with exogenous ligands

通过外源配体调节 RNA 折叠来调节 RNA 功能

• 批准号: 9119049
• 负责人: Nathan Baird
• 金额: $ 24.37万
• 依托单位: UNIVERSITY OF THE SCIENCES PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9119049
• 关键词: Advisory Committees; Affect; Area; Award; Bacterial Genes; Binding; Biochemical; Biological Assay; Biological Models; Biological Process; Biology; Biophysics; Calorimetry; Career Mobility; Career Transition Award; Catalytic RNA; Cell physiology; Cells; Charge; Chemicals; Collaborations; Complex; Development; Divalent Cations; Doctor of Philosophy; Drug Targeting; Education; Educational process of instructing; Effectiveness; Environment; Exposure to; Extramural Activities; Fluorescence; Fluorescence Resonance Energy Transfer; Genes; Genetic Transcription; Glutamine; Glycine; Goals; HIV-1; Head; Human; Hydrogen Bonding; Laboratories; Laboratory Research; Lead; Libraries; Ligand Binding; Ligands; Measures; Mediating; Mentors; Methodology; Methods; MicroRNAs; Modeling; Molecular; Monitor; Morphologic artifacts; Mutation; National Heart, Lung, and Blood Institute; Oncogenic; Pathway interactions; Phase; Positioning Attribute; Postdoctoral Fellow; Process; Professional Competence; Property; Qualifying; RNA; RNA Binding; RNA Folding; RNA Interference; Reading; Regulator Genes; Reporter; Research; Research Personnel; Resources; Roentgen Rays; Role; Scientist; Shapes; Site; Structure; Structure-Activity Relationship; Testing; Titrations; Training; Training Activity; Training and Education; Transcript; Translational Research; United States National Institutes of Health; Universities; Ursidae Family; Viral Genome; Virulence; Virus Replication; Work; X-Ray Crystallography; assay development; base; career; career development; design; drug development; fluorophore; high throughput screening; in vivo; innovation; instrumentation; laboratory equipment; novel; novel therapeutics; nucleocytoplasmic transport; pathogenic bacteria; programs; research and development; research study; screening; small molecule; small molecule libraries; success; synthetic biology; therapeutic target; tool; transcription termination

DESCRIPTION (provided by applicant): The goal of this application is to establish a focused training plan to successfully advance my research and career goals during the award period. The details provided in this proposal include specific action steps for me to gain training in career skills that I have had little exposure to that include, for example, effective grantsmanship mentoring, and laboratory management. Furthermore, to realize my research goals, I am in need of training in new scientific fields, those of high-throughput screening assays and chemical biology. To that end, I have recently established a collaboration with the laboratory of Dr. James Inglese, director of the Assay Development and Screening Technologies Laboratory in the National Center for Advancing Translational Sciences (NCATS), who will serve as my co-mentor as described below. These proposed training activities are necessary to prepare me for a successful transition to an independent investigator position. All aspects of my training will be supported by an Advisory Committee comprised of five leading scientists with distinct scientific and career expertise in both intramural and extramural research. The goal of my research program is to develop an innovative research plan aimed at leveraging the chemical diversity of high-throughput screening (HTS) libraries to explore the modulation of RNA structure by exogenous ligands, a novel methodology I refer to as HTS-MoRSEL. By targeting RNA folding directly, rather than binding, small molecule ligands will be identified that control the folding/function of nearly any structured RNA. This high-impact research program has broad implications for probing structure-function relationships within the expanding field of RNA biology. During the award period, I will identify ligands modulating the folding and function of several RNAs of biomedical significance. For example, I will examine RNAs that regulate genes responsible for viral replication or virulence in pathogenic bacteria. Additionally, I will apply tis methodology to identify ligands capable of modulating the structure of a human oncogenic microRNA polycistron; the structure and folding of this large RNA transcript are responsible for autoregulating its processing by the RNAi pathway machinery. Together, my results will present a new paradigm for the design of therapeutics targeting RNA and the development of synthetic biology tools. While I have demonstrated exceptional research success both in my Ph.D. and post-doctoral studies, the new direction of my proposed research plan necessitates additional training that will build on my existing training. Specifically, only having been introduced recentl to HTS chemical biology methodologies, I am working to establish my proficiency employing this approach. Given the prominence of these assays in my proposed research program, I am in need of focused training in this field during the mentored phase of the award period. I recently established a collaboration with Dr. James Inglese, who has agreed to serve as co-mentor during this award. I will receive focused training in his laboratory towards the development of appropriate HTS assays for my RNA folding studies. Under this Career Transition Award I will also receive necessary additional training in the lab of my mentor, Dr. Ferr�-D'Amar� (RNA Biophysics and Cellular Physiology, NHLBI) for the molecular and atomistic characterization studies of RNA-ligand interactions identified in my HTS studies. The existing state-of-the-art instrumentation resources within these labs are more than sufficient to complete all aspects of this proposed research program. Both Dr. Ferr�-D'Amar� and Dr. Inglese will work together to provide me with necessary career development training for establishing an independent research lab. Uniquely, following this training I will be aptly qualified to perform both HTS assay and molecular characterization of RNA-ligand interactions. Together, these combined approaches will make me a highly qualified candidate to lead an independent research laboratory. The environment within the NHLBI, NCATS, and NIH are exceptionally suited for the development of all aspects of this award application. Along with the scientific resources, the NIH offers extensive career development programs for post-doctoral researchers (e.g. the Career Advancement Toolkit Track) through the Office of Intramural Education and Training, headed by Dr. Sharon Milgram. Furthermore, Dr. Herbert Geller, director of the NHLBI Office of Education, offers many complementary resources and will provide me with individualized career development guidance. In summary, under the NHLBI Career Transition Award, I will receive specific training in research and career development areas in which I have had little exposure to date including, but not limited to, HTS assay development, mentoring, teaching, and lab management. Subsequent to this training I will be excellently positioned to achieve my career goal of establishing a high-impact, independent research laboratory at an academic university.

描述（由应用程序提供）：本申请的目的是建立一个重点的培训计划，以在奖励期间成功地推进我的研究和职业目标。本提案中提供的细节包括我在职业技能上获得培训的具体行动步骤，而这些培训几乎没有接触到，例如，例如有效的授予技巧和实验室管理。此外，为了实现我的研究目标，我需要在新科学领域，高通量筛查评估和化学生物学的培训。为此，我最近与国家前进转化科学中心（NCATS）的测定开发和筛查技术实验室主任詹姆斯·英格莱斯博士建立了合作，该实验室将作为我的同事，如下所述。这些提议的培训活动是为了使我成功过渡到独立研究者职位的准备工作。我的培训的所有方面都将得到一个咨询委员会的支持，该咨询委员会在壁内和壁外研究中完成了五位具有独特的科学和职业专业知识的主要科学家。我的研究计划的目的是制定旨在利用高通量筛选（HTS）库的化学多样性的创新研究计划，以通过外源配体探索RNA结构的调节，这是我称为HTS-Morsel的一种新方法。通过直接靶向RNA折叠而不是结合，将确定小分子配体控制几乎所有结构化RNA的折叠/功能。这个高影响力的研究计划对RNA生物学扩展领域内的探测结构 - 功能关系的关系具有广泛的影响。在奖励期间，我将确定调节几种生物医学意义RNA的折叠和功能的配体。例如，我将检查调节致病细菌病毒复制或病毒的基因的RNA。此外，我将采用TIS方法来识别能够调节人致癌的MicroRNA多层神的结构的配体；该大RNA转录本的结构和折叠负责通过RNAi途径机械自动调节其处理。总之，我的结果将为靶向RNA的理论设计和合成生物学工具的开发提供一个新的范式。虽然我在博士学位都取得了出色的研究成功。以及博士后研究，我提议的研究计划的新方向必要的其他培训将以我现有的培训为基础。具体而言，仅引入了HTS化学生物学方法的近期，我才努力通过这种方法来确定自己的能力。鉴于我在拟议的研究计划中对这些评估的承诺，我需要在奖励期的修订阶段进行该领域的重点培训。我最近与詹姆斯·英格莱斯（James Inglese）博士建立了合作，后者同意在此奖项中担任联合学。我将在他的实验室接受重点的培训，以开发适当的HTS分析以进行RNA折叠研究。根据该职业过渡奖，我还将在我的心态实验室中获得必要的额外培训。这些实验室内的现有最新仪器资源足以完成该建议的研究计划的所有方面。 Ferr博士‥-D'amar‥博士和Inglese博士都将共同努力，为我提供必要的职业发展培训，以建立一个独立的研究实验室。独特的是，在此训练之后，我将有资格同时执行RNA - 配体相互作用的HTS分析和分子表征。这些合并的方法在一起将使我成为领导独立研究实验室的高素质候选人。 NHLBI，NCAT和NIH中的环境非常适合开发本奖项申请的各个方面。与科学资源一起，NIH通过莎伦·米尔格拉姆（Sharon Milgram）博士领导，为博士后研究人员提供广泛的职业发展计划（例如，职业发展工具包轨道）。此外，NHLBI教育办公室主任赫伯特·盖勒（Herbert Geller）博士提供了许多完整的资源，并将在摘要中为我提供，根据NHLBI职业过渡奖，我将获得有关我在研究和职业发展领域的特定培训，在我的研究和职业发展领域中，我几乎不包括迄今为止，包括，不限于HTS评估，心理学教学，心理学教学和实验室管理。在这项培训之后，我将在一所学术大学建立高影响力，独立的研究实验室的职业目标。