Inducible Tissue-Specific Transgene Expression in Large Animal Biomedical Models

描述（由申请人提供）：本研究提案的总体目标是开发可诱导的、组织特异性表达的转基因，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，并且在啮齿动物模型中如此成功的技术在大型动物中的有限应用需要替代方法。本提案的具体目标是开发有效的组织特异性的、体外诱导表达的转基因，然后利用重组慢病毒载体将这些转基因递送到猪受精卵中，并在所产生的后代中评估表达平台。这些实验的靶基因将是硬脂酰-CoA去饱和酶（SCD-1），并且将用作检查体内基因表达的局部控制的有效且高效的模型。尽管可以利用任何数量的基因，但已经证明SCD-1在脂质生物合成中具有重要作用，并且其组织特异性表达在维持体内平衡中是重要的。因此，由此产生的动物不仅将证明该技术平台的有效性，而且还将以组织特异性方式作为脂质代谢失败的有用模型。该技术的开发和该基因靶标的利用对于研究诸如肥胖症、胰岛素抵抗和相关代谢紊乱以及肌肉和肝脏脂肪变性的疾病至关重要。最重要的是，该项目的成功完成将导致技术进步，有助于在许多大型动物物种中产生各种各样的生物医学模型，可能针对大量基因，从而使许多NIH研究所和中心受益。

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.