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是多谷氨酰胺（polyq）蛋白，可作为染色质结合因子和基因复制品起作用。在近年来，在多种病理生理过程中发现了共生1 控制APP淀粉样蛋白生成裂解，以调节海马中神经前体增殖。我们的小组还强调了一种新型的ataxin-1靶向B细胞反应的免疫调节功能中枢神经系统（CNS）自身免疫。该实验证据增加了一层复杂性进行ataxin-1研究，主要集中于脊椎动物共济失调类型（SCA1）由polyq膨胀突变引起的神经退行性疾病。重要的是，所有的努力到目前为止，ataxin-1功能的特征仅在全球ataxin-1 null小鼠上救出。但是，无处不在共生1和许多组织的复杂细胞结构的表达使系统复杂化在不同的细胞类型和发育阶段，由Ataxin-1控制的所有分子机制分析。要填补此空白，我们建议使用二进制CRE/LOX开发条件性ataxin-1敲除鼠标线系统。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作用任何靶标细胞型中的ataxin-1。在特定目标1中，我们将使用CRISPR/CAS9技术介绍两个小鼠基因组中的LOXP位点，侧翼是编码ATXN1基因的Ataxin-1外显子。在特定的目标2中，我们将繁殖新生成的Flox Atxn1小鼠系列，并用CD19-CRE驱动线将ataxin-1烧在B中细胞室。然后，我们将诱导多发性硬化模型实验性自身免疫性脑脊髓炎（EAE）在条件ATXN1基因敲除小鼠中，以研究B细胞中ataxin-1表达的贡献自身免疫性脱髓鞘。总共提出的目标持有生成新的研究工具的能力这将大大提高对ataxin-1生物学的当前理解，并具有重要的含义和几种神经退行性疾病的课程，包括多发性硬化症和阿尔茨海默氏病。