Redesign Rat Model for ALS Research

重新设计用于 ALS 研究的大鼠模型

基本信息

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
项目类别：
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

项目摘要

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（由申请人提供）：某些基因的基因突变会导致家族性肌萎缩性侧索硬化症（ALS）。为了理解疾病突变的机制，一个关键的步骤是为研究创造理想的模型。尽管细胞模型对于测试疾病机制的某些方面很重要，但动物模型对于揭示复杂神经变性的机制至关重要，因为体外研究的发现并不总是转化为活动物。当致病性突变表现出显性性状时，动物模型通常是通过将转基因插入宿主基因组而产生的，从而产生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，即使两条线携带相同的转基因副本，也会因线对线而有所不同。遗传模型的最佳控制是表达疾病基因的野生型形式但在突变转基因菌株中未观察到的表型的转基因菌株。不太可能获得两条线以相似的水平和模式表达野生型和突变蛋白。真实的机制通常被传统转基因的缺点所掩盖。为了避免基因插入突变并统一单个线之间的位置效应，我们将在预定的基因间基因座上插入单个转基因的副本，并将其与TET诱导的系统结合起来，以扩大转基因的表达，以便在大鼠的寿命内诱导可见的表型。使用这些新型大鼠，我们将在系统水平上检查HNRNPA1发病机理的机制。我们将通过将表型表达与在生理水平及其内在模式中表达疾病基因的敲击大鼠中观察到的表型表达进行比较，从而验证这种新型转基因诱导中这种新型转基因方法的功效。