RNA-based tools for developmental and regenerative biology

用于发育和再生生物学的基于 RNA 的工具

• 批准号: 10287427
• 负责人: Maureen Hope McKeague
• 金额: $ 19.21万
• 依托单位: UNIVERSITY OF WYOMING
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10287427
• 关键词: Address; Animal Model; Benchmarking; Biological; Biomedical Research; Cell model; Cells; Chemicals; Complement; Complex; Development; Developmental Biology; Dose; Embryo; Event; Fluorescence Microscopy; Gene Expression; Genes; Genetic; Genetic Screening; Genetic Transcription; Goals; Guanine; Interdisciplinary Study; Laboratories; Leucovorin; Light; Mammalian Cell; Methods; Modeling; Molecular; Natural regeneration; Optics; Organism; Pattern; Periodicity; Permeability; Photochemistry; Photoreceptors; Positioning Attribute; Process; Proteins; RNA; Regenerative capacity; Reporter Genes; Research; Signal Pathway; System; Techniques; Technology; Testing; Tetracyclines; Theophylline; Time; Tissues; Translations; Vertebrates; Whole Organism; Work; Xanthines; Zebrafish; base; dosage; gene function; genetic manipulation; genome sequencing; innovation; insight; novel; novel strategies; optogenetics; programs; prototype; regenerative; regenerative biology; sensor; small molecule; spatiotemporal; technology development; tool

Project Summary RNA-based technologies have enabled perturbation and observation of gene function in multicellular organisms, resulting in major discoveries in biomedical research. However, gaining mechanistic insights into developmental and regenerative processes requires the manipulation of gene functions with spatial and temporal precision, a task that remains a major challenge in vertebrate animal models. The overall goal of this exploratory technology development proposal is to develop an innovative and widely applicable molecular toolset, which will enable control of gene expression with spatial and temporal precision. Specifically, we will introduce a new class of versatile RNA-based genetic conditional switches to regulate translation within the developing zebrafish. In our first aim, we will develop RNA switches that are controlled through non-invasive administration of non-toxic small molecules. These chemically-induced switches enable dynamic, titratable, and conditional control of a target gene. We will establish a rapid mammalian- based prototyping platform to prioritize ideal switch candidates for development in the zebrafish model. In our second aim, we will establish a conceptually new optogenetic switch that can be turned on and off using blue light. These novel tools will be developed in zebrafish and benchmarked against existing optogenetic tools including light-activated transcriptional tools. The successful execution of this project will provide a streamlined pipeline to develop new RNA switches that respond to a diversity of orthogonal non-invasive inputs. Given the species-independent machinery, the toolkit of RNA switches developed are likely to be broadly applicable to animal models. By specifically focusing the development of the switches in zebrafish, we will we will contribute a new approach to reprogram and interrogate developmental and regenerative biology.

项目概要 基于 RNA 的技术使得多细胞基因功能的扰动和观察成为可能 生物体，导致生物医​​学研究的重大发现。然而，获得机械见解 发育和再生过程需要对基因功能进行空间和 时间精度，这项任务仍然是脊椎动物模型中的一个主要挑战。总体目标为 这项探索性技术开发建议旨在开发一种创新且广泛适用的技术 分子工具集，它将能够以空间和时间精度控制基因表达。 具体来说，我们将引入一类新的基于 RNA 的多功能遗传条件开关来调节 发育中的斑马鱼内的翻译。我们的第一个目标是开发受控的 RNA 开关 通过无毒小分子的非侵入性给药。这些化学感应开关 实现对目标基因的动态、可滴定和条件控制。我们将建立一个快速的哺乳动物- 基于原型平台，优先考虑在斑马鱼模型中开发的理想开关候选者。在 我们的第二个目标是，我们将建立一种概念上新的光遗传学开关，可以使用它来打开和关闭 蓝光。这些新颖的工具将在斑马鱼中开发，并以现有的光遗传学为基准 工具包括光激活转录工具。该项目的成功实施将为 简化流程以开发新的 RNA 开关，以响应各种正交非侵入性 输入。考虑到与物种无关的机制，开发的 RNA 开关工具包很可能是 广泛适用于动物模型。通过特别关注斑马鱼开关的开发， 我们将贡献一种新的方法来重新编程和询问发育和再生 生物学。