Regulating RNA function by modulating RNA folding with exogenous ligands

描述（由申请人提供）：本申请的目的是建立一个有针对性的培训计划，以成功推进我在奖励期间的研究和职业目标。该提案中提供的细节包括我获得职业技能培训的具体行动步骤，这些技能我几乎没有接触过，例如，有效的资助指导和实验室管理。此外，为了实现我的研究目标，我需要在新的科学领域，高通量筛选分析和化学生物学方面进行培训。为此，我最近与国家促进转化科学中心（NCATS）检测开发和筛选技术实验室主任James Inglese博士的实验室建立了合作关系，他将担任我的共同导师，如下所述。这些拟议的培训活动对我成功过渡到独立调查员的职位是必要的。我的培训的所有方面都将得到一个咨询委员会的支持，该委员会由五名在校内外研究方面具有独特科学和职业专长的顶尖科学家组成。我的研究计划的目标是制定一个创新的研究计划，旨在利用高通量筛选（HTS）文库的化学多样性来探索外源性配体对RNA结构的调节，这是一种我称之为HTS- morsel的新方法。通过直接靶向RNA折叠，而不是结合，将确定控制几乎任何结构RNA的折叠/功能的小分子配体。这个高影响力的研究项目对探索RNA生物学不断扩大的领域中的结构-功能关系具有广泛的意义。在获奖期间，我将识别调节几种具有生物医学意义的rna的折叠和功能的配体。例如，我将研究在致病菌中调节负责病毒复制或毒力的基因的rna。此外，我将应用这种方法来鉴定能够调节人类致癌微rna多顺子结构的配体；这种大RNA转录物的结构和折叠负责通过RNAi途径机制自动调节其加工。总之，我的研究结果将为设计靶向RNA的治疗方法和开发合成生物学工具提供一个新的范例。虽然我在博士和博士后研究中都取得了非凡的研究成功，但我提出的研究计划的新方向需要在现有培训的基础上进行额外的培训。具体地说，我最近才接触到HTS化学生物学方法，我正在努力建立我使用这种方法的熟练程度。鉴于这些分析在我的研究计划中的重要性，我需要在奖励期间的指导阶段在这一领域进行重点培训。我最近与James Inglese博士建立了合作关系，他同意在本次颁奖期间担任联合导师。我将在他的实验室接受重点培训，为我的RNA折叠研究开发合适的HTS检测方法。在这个职业过渡奖下，我还将在我的导师Ferr ' d 'Amar博士（RNA生物物理学和细胞生理学，NHLBI）的实验室接受必要的额外培训，以进行我在HTS研究中发现的RNA-配体相互作用的分子和原子表征研究。这些实验室现有的最先进的仪器资源足以完成这个拟议的研究计划的各个方面。Dr. Ferr -D - amar和Dr. Inglese将共同合作，为我建立独立的研究实验室提供必要的职业发展培训。独特的是，在这次培训之后，我将有资格进行HTS分析和rna -配体相互作用的分子表征。总之，这些结合起来的方法将使我成为领导一个独立研究实验室的非常合格的候选人。NHLBI、NCATS和NIH内部的环境非常适合该奖项申请的各个方面的开发。除了科学资源，NIH还通过Sharon Milgram博士领导的校内教育和培训办公室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跟踪）。此外，NHLBI教育办公室主任Herbert Geller博士提供了许多补充资源，并将为我提供个性化的职业发展指导。总而言之，在NHLBI职业转型奖的资助下，我将在研究和职业发展领域接受具体的培训，这些领域是我迄今为止几乎没有接触过的，包括但不限于HTS检测开发、指导、教学和实验室管理。通过这次培训，我将能够很好地实现我的职业目标，在一所学术大学建立一个具有高影响力的独立研究实验室。

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.