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.

项目摘要 基于RNA的技术已经使得能够在多细胞中干扰和观察基因功能。 生物体，导致生物医学研究的重大发现。然而，获得机械的见解， 进入发育和再生过程需要操纵基因功能， 时间精度，这是脊椎动物模型中的一个主要挑战。的总目标 这一探索性的技术发展建议是开发一种创新的和广泛适用的 分子工具集，这将使控制基因表达的空间和时间精度。 具体来说，我们将介绍一类新的多功能RNA为基础的遗传条件开关，以调节 在发育中的斑马鱼中进行翻译。在我们的第一个目标中，我们将开发一种RNA开关， 通过非侵入性的无毒小分子给药。这些化学诱导的开关 能够动态、可滴定和有条件地控制靶基因。我们将建立一个快速的哺乳动物- 基于原型平台，优先考虑理想的开关候选人在斑马鱼模型的发展。在 我们的第二个目标是，我们将建立一个概念上新的光遗传学开关，可以打开和关闭， 蓝光这些新工具将在斑马鱼中开发，并以现有的光遗传学为基准。 包括光激活转录工具的工具。该项目的成功执行将提供 流线型的管道，以开发新的RNA开关，响应于正交的非侵入性的多样性 输入。鉴于这种不依赖于物种的机制，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中开关的开发， 我们将提供一种新的方法来重新编程和询问发育和再生 生物学