Inducible Tissue-Specific Transgene Expression in Large Animal Biomedical Models

描述(由申请人提供)：这项研究计划的总体目标是开发可诱导的、组织特异性的转基因表达，以产生人类疾病的大型动物模型。迫切需要将功能基因组学扩展到非啮齿动物模型，而在啮齿动物模型中取得如此成功的技术在大型动物中的有限应用需要另一种方法。该方案的具体目标是在体外建立有效的组织特异性、可诱导的转基因表达，然后利用重组慢病毒载体将这些转基因导入猪受精卵中，并对所产生的后代的表达平台进行评估。这些实验的目标基因将是硬脂酰辅酶A去饱和酶(SCD-1)，并将成为检查体内基因表达的局部调控的有效和高效的模型。虽然可以利用许多基因，但SCD-1已被证明在脂质生物合成中具有重要作用，其组织特异性表达在维持体内平衡方面具有重要意义。因此，由此产生的动物不仅将展示技术平台的有效性，而且还将以组织特异性的方式作为脂肪代谢失败的有用模型。这项技术的发展和对该基因靶点的利用是研究肥胖、胰岛素抵抗和相关代谢紊乱以及肌肉和肝脏脂肪变性等疾病的核心。最重要的是，该项目的成功完成将带来有助于在众多大型动物物种中生产各种生物医学模型的技术进步，潜在地针对大量的基因，从而使许多NIH研究所和中心受益。 公共卫生相关性(由申请人提供)：开发治疗人类疾病的疗法需要对相关动物模型中的基因功能进行广泛调查。当自然产生的动物模型不存在时，生产模仿人类疾病的基因工程动物是必要的。我们的项目将开发可靠地诱导组织特异性基因表达的技术，以便可以在非啮齿动物物种中模拟疾病状态。

DESCRIPTION (provided by applicant): The overall goal of this research proposal is to develop inducible, tissue-specific expression of transgenes to produce large animal models of human disease. There is a critical need to expand functional genomics to non-rodent models and the limited application of technologies, which have been so successful in rodent models, in large animals requires an alternate approach. The specific objective of this proposal is to develop effective tissue-specific, inducible expression of transgenes in vitro, then utilize recombinant lentiviral vectors to deliver these transgenes into porcine zygotes and evaluate the expression platforms in resulting offspring. The target gene for these experiments will be Stearoyl-CoA desaturase (SCD-1) and will serve as an effective and efficient model for examining local control of gene expression in vivo. Although any number of genes could be utilized, SCD-1 has been documented to have an important role in lipid biosynthesis and its tissue specific expression is important in maintaining homeostasis. Therefore, the resulting animals will not only demonstrate the effectiveness of the technology platform, but also serve as a useful model of failed lipid metabolism in a tissue-specific manner. Development of this technology, and utilization of this gene target is central to investigation of diseases such as obesity, insulin resistance and associated metabolic disorders, as well as, muscle and hepatic steatosis. Most importantly, successful completion of this project will result in technological advancements useful to produce a wide variety of biomedical models in numerous large animal species, potentially targeting a huge range of genes, thus benefiting many of the NIH Institutes and Centers. PUBLIC HEALTH RELEVANCE (provided by applicant): Developing therapies to treat human disease requires extensive investigation of gene function in relevant animal models. When naturally occurring animal models do not exist, production of genetically engineered animals that mimic human disease is necessary. Our project will develop the technology to reliably induce tissue-specific expression of genes so that disease states can be mimicked in non-rodent species.