Development of a conditional ataxin-1 knockout mouse line

条件性ataxin-1基因敲除小鼠品系的开发

基本信息

批准号：
10642313
负责人：
Alessandro Didonna
金额：
$ 7.55万
依托单位：
EAST CAROLINA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-02-15 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10642313
关键词：
Ablation Address Adopted Adult Alzheimer&apos s Disease Antigen Presentation Autoimmune Autoimmune Responses B-Cell Activation B-Lymphocytes Binding Biological Process Biology Breeding CNS autoimmunity CRISPR/Cas technology Cell Compartmentation Cell Nucleus Cell Proliferation Cell physiology Cells Central Nervous System Chromatin Communities Complement Complex Core Facility Cre driver Cre-LoxP Cytosol Demyelinations Development Discrimination Disease Disease Progression Dissection Embryo Enterobacteria phage P1 Cre recombinase Enzyme-Linked Immunosorbent Assay Epithelium Exons Experimental Autoimmune Encephalomyelitis Experimental Models Female Flow Cytometry Funding Gene Targeting Genes Grant Guide RNA Health Heterogeneity Hippocampus Immune Immune system Immunoglobulins Immunohistochemistry Immunophenotyping Implant Inflammation Knock-out Knockout Mice Laboratories LoxP-flanked allele Measures Mediating Mesenchymal Methods Microinjections Modeling Molecular Multiple Sclerosis Mus Mutate Mutation Nervous System Neurodegenerative Disorders Neurons North Carolina Nuclear Organism Outsourcing Pathologic Phenotype Physiological Population Process Proliferating Proteins Purkinje Cells Regulation Research Resources Role Sampling Serum Signal Transduction Site Spinocerebellar Ataxias System Technology Tissues Transcriptional Regulation Type 1 Spinocerebellar Ataxia Universities Work ataxin-1 axon injury beta-site APP cleaving enzyme 1 cancer cell cell type conditional knockout cytokine design essays gain of function mutation gene correction genome editing histological stains immunoregulation in vivo Model intercellular communication interest loss of function male mouse genome mouse model nerve stem cell neurogenesis novel polyglutamine programs promoter response screening single-cell RNA sequencing tool tumorigenesis vector

项目摘要

Summary Ataxin-1 is polyglutamine (polyQ) protein that operates as a chromatin-binding factor and gene repressor. In recent years, ataxin-1 has been found implicated in multiple pathophysiological processes ranging from the control of APP amyloidogenic cleavage to the modulation of neural precursor proliferation in the hippocampus. Our group has also highlighted a novel immunomodulatory function for ataxin-1 targeting B cell responses in central nervous system (CNS) autoimmunity. This experimental evidence adds an additional layer of complexity to ataxin-1 research, which has been mainly focused on spinocerebellar ataxia type (SCA1)—a neurodegenerative disorder caused by polyQ expansion mutations in ataxin-1. Importantly, all the efforts carried out so far to characterize ataxin-1 function have relied only on global ataxin-1 null mice. However, the ubiquitous expression of ataxin-1 and the complex cytoarchitecture of many tissues have complicated the systematic analysis of all the molecular mechanisms controlled by ataxin-1 in different cell types and developmental stages. To fill this gap, here we propose to develop a conditional ataxin-1 knockout mouse line using the binary Cre/Lox system. This novel in vivo model will be instrumental to dissect the cell-autonomous and cell-mediated effects of ataxin-1 in any target cytotype. In Specific Aim 1, we will employ CRISPR/Cas9 technology to introduce two LoxP sites in the mouse genome, flanking one exon of the ataxin-1 encoding Atxn1 gene. In Specific Aim 2, we will breed the newly generated floxed Atxn1 mouse line with a Cd19-Cre driver line to ablate ataxin-1 in the B cell compartment. We will then induce the multiple sclerosis model experimental autoimmune encephalomyelitis (EAE) in the conditional Atxn1 knockout mice, in order to study the contribution of ataxin-1 expression in B cells to autoimmune demyelination. Altogether, the proposed aims hold the power to generate a new research tool that will substantially advance the current understanding of ataxin-1 biology, with important implications and lessons for several neurodegenerative diseases including multiple sclerosis and Alzheimer's disease.

概括 Ataxin-1 是一种聚谷氨酰胺 (polyQ) 蛋白，可作为染色质结合因子和基因阻遏蛋白。近年来，人们发现ataxin-1与多种病理生理过程有关，从控制 APP 淀粉样蛋白生成裂解来调节海马神经前体增殖。我们的小组还强调了 ataxin-1 靶向 B 细胞反应的新型免疫调节功能中枢神经系统（CNS）自身免疫的实验证据增加了一层额外的复杂性。 ataxin-1 研究主要集中在脊髓小脑性共济失调型 (SCA1) 重要的是，所有的努力都取得了成果。迄今为止，对 ataxin-1 功能的表征仅依赖于全局 ataxin-1 缺失小鼠。 ataxin-1 的表达和许多组织的复杂细胞结构使系统的复杂化分析不同细胞类型和发育阶段中ataxin-1控制的所有分子机制。为了填补这一空白，我们建议使用二元 Cre/Lox 开发条件 ataxin-1 基因敲除小鼠系这种新颖的体内模型将有助于剖析细胞自主和细胞介导的效应。在特定目标1中，我们将利用CRISPR/Cas9技术引入两种靶细胞型中的ataxin-1。小鼠基因组中的 LoxP 位点，位于编码 Atxn1 基因的 ataxin-1 的一个外显子两侧。将用 Cd19-Cre 驱动系培育新生成的 floxed Atxn1 小鼠系，以消除 B 中的 ataxin-1 然后我们将诱导多发性硬化症实验性自身免疫性脑脊髓炎模型。 (EAE) 在条件 Atxn1 敲除小鼠中，以研究 ataxin-1 表达在 B 细胞中的贡献总而言之，所提出的目标有能力产生一种新的研究工具。这将大大推进目前对 ataxin-1 生物学的理解，具有重要意义和包括多发性硬化症和阿尔茨海默病在内的多种神经退行性疾病的教训。