Illuminating cellular dark matter through the development of novel chemical tools

通过开发新型化学工具照亮细胞暗物质

• 批准号: 10581981
• 负责人: Neal Krishna Devaraj
• 金额: $ 8.98万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10581981
• 关键词: Affect; Amyotrophic Lateral Sclerosis; Biological; Biology; C9ORF72; Cell physiology; Cells; Cellular biology; Chemicals; Development; Disease; Genetic; Goals; Image; In Situ; Knowledge; Laboratories; Lead; Lipids; Modification; Molecular; Nucleotides; Pathogenicity; Pathway interactions; Permeability; Play; Proteins; RNA; Research; Role; Site; Sphingolipids; Technology; Therapeutic Intervention; Vision; cell behavior; cytotoxic; dark matter; human disease; improved; interest; novel; programs; protein complex; recruit; success; tool

PROJECT SUMMARY There is a vast repertoire of species within cells for which we have a poor understanding of their function and biomolecular interactions. These species can be referred to as the “dark matter” of biology, as their mechanism of action is hidden from conventional observation. Our laboratory seeks to illuminate the function of cellular “dark matter” through the development of new chemical technology. The proposed research program will pursue two major research thrusts. First, we plan to develop tools for site-specific RNA modification, and apply these tools for the manipulation, imaging, and isolation of disease relevant RNA protein complexes. We will create tools for use in live cells and develop the ability to covalently recruit proteins to RNA, forming RNA-protein macromolecular conjugates. The technology will be applied to study RNAs implicated in disease. Specifically, we are interested in characterizing the pathways of pathogenicity for the C9orf72 nucleotide repeat expansion RNA, which is thought to play a major role in genetic amyotrophic lateral sclerosis (ALS). In the second thrust, we will carry out the in situ synthesis of lipid species within living cells, with the goal of uncovering the molecular mechanism by which enigmatic lipid species affect cell behavior. We plan to develop approaches enabling the selective and bioorthogonal delivery of sphingolipids to living cells. Building upon technology previously developed in our lab, we will deliver cell permeable lipid precursors which will spontaneously assemble into functional lipids within the cell. Leveraging this approach, we will create photoaffinity probes for the pulldown of sphingolipid-interacting proteins, with the goal of elucidating the protein partners of the non-canonical deoxysphingolipid 1-deoxydihydroceramide, which is cytotoxic and implicated in several diseases. Realization of our research program goals would improve our knowledge of cell biology and lead to the development of new tools for interrogating RNA and lipid species. Our long-term vision is to create and apply technology that enables improved mechanistic understanding of biomolecular interactions, leading to an increased understanding of human disease, and accelerating the development of possible therapeutic interventions.

项目摘要 细胞内有广泛的物种曲目，我们对它们的功能的了解不足 生物分子相互作用。这些物种可以称为生物学的“暗物质”，作为其机制 动作不受传统观察的隐藏。我们的实验室试图照亮细胞的功能“黑暗 通过开发新的化学技术。拟议的研究计划将追求两个 主要的研究推力。首先，我们计划开发用于特定网站的RNA修改工具，并应用这些工具 用于操作，成像和孤立疾病相关的RNA蛋白复合物。我们将为 在活细胞中使用，并发展能够共价募集蛋白质到RNA的能力，从而形成RNA蛋白 大分子结合物。该技术将用于研究疾病实施的RNA。具体来说， 我们有兴趣表征C9orf72核苷酸重复扩张的致病性途径 RNA被认为在遗传肌萎缩性侧索硬化症（ALS）中起主要作用。在第二个推力中， 我们将在活细胞内进行脂质物种的原位合成，目的是揭示分子 神秘脂质物种影响细胞行为的机制。我们计划开发方法，使 鞘脂递送到活细胞的选择性和生物共振递送。以前以技术为基础 在我们的实验室中开发的，我们将提供可渗透的脂质前体，这些脂质前体将赞助 - 细胞内的功能性脂质。利用这种方法，我们将为下拉的光性问题创建 鞘脂相互作用的蛋白质，目的是阐明非典型的蛋白质伴侣 脱氧胆脂1-脱氧二氢可酰胺，在几种疾病中是细胞毒性的。实现 我们的研究计划目标将提高我们对细胞生物学的了解，并导致新的发展 询问RNA和脂质物种的工具。我们的长期愿景是创建和应用能够启用的技术 改善了对生物分子相互作用的机械理解，从而增加了对 人类疾病，并加速可能的治疗干预措施。