Control of gene expression via RNA-targeted covalent probes

通过 RNA 靶向共价探针控制基因表达

• 批准号: 10607121
• 负责人: Alexandra Khitun
• 金额: $ 7.18万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10607121
• 关键词: Address; Affect; Bar Codes; Binding; Binding Sites; Bioinformatics; Biological; Biological Process; Biology; Categories; Cells; Chemicals; Chemistry; Complex; Degradation Pathway; Derivation procedure; Fluorescent Probes; Foundations; Gene Expression; Goals; Guanine; Health; Human; Libraries; Ligand Binding; Ligands; Link; Location; Mass Spectrum Analysis; Mechlorethamine; Messenger RNA; Modification; Molecular; Molecular Biology; Nucleotides; Oligonucleotides; Open Reading Frames; Organic Chemistry; Positioning Attribute; Proteins; Proteomics; Quality Control; RNA; RNA Binding; RNA Sequences; RNA library; RNA-targeting therapy; Reaction; Regulation; Research; Research Personnel; Research Project Grants; Research Proposals; Resolution; Reverse Transcription; Ribosomes; Role; Site; Structure; System; T47D; Technology; Therapeutic; Training; Transcript; Translations; Work; adduct; bioinformatics pipeline; biological systems; career; computational chemistry; computer science; design; drug development; drug discovery; drug-like compound; druggable target; experience; experimental study; innovation; mRNA Stability; multidisciplinary; new technology; novel; protein function; rational design; scaffold; screening; skills; small molecule; small molecule libraries; targeted treatment; technology development; technology platform; therapeutic development; therapeutic target; training project; transcriptome; transcriptome sequencing; transcriptomics

Project Summary RNA is now recognized as a compelling therapeutic target because it lies upstream of all protein function and has cellular roles that go beyond protein translation. Similar to proteins, RNA molecules can fold to create complex tertiary structures that form binding pockets capable of specific small-molecule engagement. However, compared to proteins, few small molecules have been rationally designed to target RNA. The central challenges to developing RNA-targeting ligands are, first, the difficulty of identifying ligandable sites on RNA and, second, the lack of direct strategies for converting ligand binding into functional perturbation of mRNA stability, translation and regulation. Both of these challenges can be addressed by a new ligand-engagement technology developed in our lab. This technology enables nucleotide-resolution capture of ligand-RNA interactions via a high-throughput, cell-based screen of a small-molecule library versus an RNA library – the cellular transcriptome. In preliminary work, the ligand-engagement technology has been validated with photo- reactive probes in bacterial cells, and shown to be modular and applicable to diverse chemistries and biological systems. The first goal of this training and research proposal is to extend the technology to human cells and to develop chemistries that yield covalent adducts with RNA specifically at sites of RNA tertiary structures. The second goal of this proposal is to characterize the effects of covalent modification on gene expression for a subset of ligandable RNAs. Completion of these aims will create a concise strategy for linking small molecule engagement with RNA to direct functional modulation of gene expression. This multi-disciplinary project emphasizes training in the fields of technology development, RNA biology, and bioinformatics. Expertise in these fields, combined with my previous experiences as a chemical biologist and proteomicist, will position me at the forefront of RNA-targeted therapeutics development. I envision using this training as a launching point for an independent career as a scientific investigator and visionary leader with a research agenda focused on developing new technologies that modulate RNA function and impact human health.

项目摘要 RNA现在被认为是一个引人注目的治疗靶点，因为它位于所有蛋白质功能的上游， 在细胞中的作用超越了蛋白质翻译。与蛋白质类似，RNA分子可以折叠， 复杂的三级结构，形成能够特异性小分子结合的结合口袋。 然而，与蛋白质相比，很少有小分子被合理地设计成靶向RNA。中央 开发RNA靶向配体的挑战首先是难以识别RNA上的可配位点 第二，缺乏将配体结合转化为mRNA功能扰动的直接策略 稳定性、翻译和调节。这两个挑战都可以通过新的配体参与来解决 我们实验室开发的技术。该技术能够实现配体-RNA的核苷酸分辨率捕获 通过小分子文库与RNA文库的高通量、基于细胞的筛选， 细胞转录组在初步工作中，配体接合技术已经通过照片验证， 反应探针在细菌细胞，并显示是模块化的，适用于不同的化学和生物 系统.这项培训和研究提案的第一个目标是将该技术扩展到人类细胞， 开发能在RNA三级结构位点特异性地与RNA产生共价加合物的化学方法。的 该建议的第二个目标是表征共价修饰对基因表达的影响， 可配体RNA的子集。这些目标的完成将创造一个简洁的策略， 与RNA接合以指导基因表达的功能调节。这个多学科的项目 强调技术开发、RNA生物学和生物信息学领域的培训。专门知识 这些领域，结合我以前作为化学生物学家和蛋白质组学家的经验，将使我定位于 处于RNA靶向治疗发展的最前沿。我设想把这次培训作为一个起点， 作为一名科学研究者和有远见的领导者的独立职业生涯，其研究议程侧重于 开发调节RNA功能和影响人类健康的新技术。