Redesign Rat Model for ALS Research

重新设计用于 ALS 研究的大鼠模型

• 批准号: 9269629
• 负责人: xugang xia
• 金额: $ 31.48万
• 依托单位: UNIVERSITY OF CENTRAL FLORIDA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9269629
• 关键词: Adult; Amyotrophic Lateral Sclerosis; Animal Model; Animals; Astrocytes; Axonal Transport; Biological Models; Cell model; Cell physiology; Cells; Chromosomes, Human, Pair 14; Complex; DNA Sequence Alteration; Disease; Disease model; Dominant Genetic Conditions; Exhibits; Gene Exchanges; Gene Targeting; Genes; Genetic; Genetic Models; Genome; Genotype; Glutamates; Heterogeneous-Nuclear Ribonucleoproteins; In Vitro; Individual; Insertion Mutation; Integrase; Knock-in; Knock-out; Link; Modeling; Motor Neurons; Mus; Mutation; Nerve Degeneration; Neurons; Pathogenesis; Pathogenicity; Pathology; Pattern; Pharmacological Treatment; Phenotype; Physiological; Positioning Attribute; Proteins; Rattus; Reproducibility; Research; Rodent; Rodent Model; Site; System; Testing; Tetracyclines; Therapeutic; Toxic effect; Transgenes; Transgenic Animals; Transgenic Mice; Transgenic Organisms; Translating; behavior test; design; disease phenotype; gene function; in vivo; inducible gene expression; mouse genome; mutant; neuron loss; novel; overexpression; performance tests; public health relevance; rat genome; resilience; superoxide dismutase 1; transgene expression

DESCRIPTION (provided by applicant): Genetic mutation in certain genes causes familial forms of amyotrophic lateral sclerosis (ALS). Toward understanding the mechanisms of a disease mutation, a critical step is creating a desirable model for research. Whereas cellular models are important for testing some aspects of disease mechanisms, animal model is essential for unraveling the mechanisms of complex neurodegeneration because findings from in vitro studies do not always translate to live animals. When a pathogenic mutation exhibits a dominant trait, animal models are often created by randomly inserting transgenes into host genome, incurring gene-insertional mutation and position effects. The levels and patterns of transgene expression largely depend on position effects and vary from line to line even when two lines carry the same copy of transgenes. The best control for genetic model is the transgenic strain that expresses wildtype form of a disease gene but develops no phenotypes observed in mutant transgenic strain. It is unlikely to obtain two lines that express wildtype and mutant proteins at similar levels and patterns. Authentic mechanisms are often obscured by the drawbacks of traditional transgenics. To avoid gene insertional mutation and unify position effect among individual lines, we will insert single copy of transgene at predetermined intergenic locus and will combine it with Tet-inducible system to amplify transgene expression such that plausible phenotypes can be induced within a rat's lifetime. Using these novel rats, we will examine the mechanisms of hnRNPA1 pathogenesis at a systematic level. We will validate the efficacy of this novel transgenic approach in disease induction by comparing phenotypic expression with those observed in the knockin rats that express disease gene at physiological levels and in its intrinsic patterns.

描述(申请人提供)：某些基因的基因突变会导致家族性肌萎缩侧索硬化症(ALS)。要了解疾病突变的机制，关键的一步是创建一个理想的研究模型。虽然细胞模型对于测试疾病机制的某些方面很重要，但动物模型对于解开复杂的神经退化机制是必不可少的，因为体外研究的结果并不总是转化为活的动物。当致病突变表现出显性特征时，通常通过在宿主基因组中随机插入转基因来建立动物模型，从而引起基因插入突变和位置效应。转基因表达的水平和模式在很大程度上取决于位置效应，甚至当两个品系携带相同的转基因拷贝时，也会因品系而异。遗传模型最好的对照是表达疾病基因野生型的转基因菌株，但在突变的转基因菌株中没有观察到的表型。不太可能获得两个以相似水平和模式表达野生型和突变型蛋白的株系。真正的机制往往被传统转基因技术的缺陷所掩盖。为了避免基因插入突变和统一个体间的位置效应，我们将在预定的基因间隔点插入转基因的单拷贝，并将其与Tet诱导系统相结合来扩增转基因表达，从而在大鼠的有生之年内诱导出合理的表型。利用这些新的大鼠，我们将在系统水平上研究hnRNPA1的发病机制。我们将通过比较在生理水平和内在模式中表达疾病基因的敲门大鼠的表型表达来验证这种新的转基因方法在疾病诱导中的有效性。