Generation of a Light Inducible Cre Transgenic Animal for KidneyResearch

用于肾脏研究的光诱导 Cre 转基因动物的产生

• 批准号: 10286292
• 负责人: John M Parant
• 金额: $ 22.28万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10286292
• 关键词: Acute; Address; Alleles; Animal Disease Models; Animal Model; Area; Biochemical; Biological Markers; Biological Models; Biosensor; Cartoons; Cell Culture Techniques; Cells; Complementary DNA; Cyst; Defect; Development; Disease; Disease Progression; Drosophila genus; Environment; Epithelial; Freedom; Future; Generations; Genetic; Genetic Recombination; Genome; Goals; Hematopoietic; Hormones; In Situ; Individual; Kidney; Knock-out; Label; Light; Loss of Heterozygosity; Methods; Modeling; Monitor; Mus; Mutation; Optics; Organism; Partner in relationship; Pathogenicity; Phase; Phenotype; Polycystic Kidney Diseases; Pre-Clinical Model; Proteins; Reporter; Research; Research Personnel; Research Project Grants; Signal Pathway; System; Tamoxifen; Temperature; Testing; Tissues; Transgenes; Transgenic Animals; Transgenic Organisms; Transplantation; Whole Organism; Zebrafish; base; cDNA Expression; cell type; conditional knockout; data access; disorder prevention; genetic manipulation; in vitro testing; interest; knockout animal; loss of function; migration; optogenetics; phase 2 testing; promoter; protein expression; recombinase-mediated cassette exchange; single cell analysis; tissue injury; tool; urologic

ABSTRACT Animal modeling of disease states and bioreporters for in situ analysis of cellular dynamics are an essential aspect of kidney, urological and hematopoietic research; however, most studies focus on genetic manipulation within the whole organism or kidney specific cell types, which does not accurately reflect disease states that are often associated with defects in individual cells or groups of cells. This discrepancy is largely due to the technical chal- lenges of being able to unrestrictedly induce Cre recombination in an optically selected group of cells within an intact organism. Therefore, we believe that a system for controlled genetic manipulation (resulting in loss of function alleles or expression of exogenous cDNAs/bioreporters) within a group of cells of a viable organism is required. This system should not alter or perturb the surrounding environment and should allow for potential systems based analysis and biomarker incorporation. Therefore, we hypothesize that transgenic lines ubiqui- tously expressing a light inducible Cre would allow for such unseen optical spatial and temporal control within a living organism. This Light Inducible Recombination (LIR) systems will be established within both the mouse and the zebrafish model organisms. Upon completion of this proposal we will have established versatile and efficient tools for selective genetic alterations that will facilitate regional and potentially single cell analysis in a multitude of research fields. We anticipate that this research will have a broad positive impact on a number areas of kidney, urological and hematopoietic research.

摘要 疾病状态的动物模型和用于原位细胞动力学分析的生物报告是一个重要方面 肾脏，泌尿和造血研究;然而，大多数研究集中在遗传操作内， 整个生物体或肾脏特异性细胞类型，这不能准确反映疾病状态， 与单个细胞或细胞组中的缺陷相关。这种差异主要是由于技术上的挑战- 本发明涉及能够在光选择的细胞组中不受限制地诱导Cre重组的方法。 完整的有机体因此，我们认为，一个控制基因操作的系统（导致基因丢失）， 功能等位基因或外源cDNA/生物报告基因的表达）在活生物体的一组细胞内是 必需的.该系统不应改变或干扰周围环境，并应考虑到潜在的 基于系统的分析和生物标记物掺入。因此，我们假设转基因株系在- 充分表达光诱导Cre将允许在光诱导的细胞内进行这种不可见的光学空间和时间控制。 活的有机体这种光诱导重组（LIR）系统将在小鼠体内建立 和斑马鱼模式生物。在完成这项建议后，我们将建立一个多功能和 选择性遗传改变的有效工具，这将有助于区域和潜在的单细胞分析， 众多的研究领域。我们预计，这项研究将对许多领域产生广泛的积极影响。 肾脏、泌尿和造血研究。