A transgenic model of ocular inflammation

描述(申请人提供)：炎症性眼病是全球范围内视力下降和失明的主要原因。葡萄膜炎的动物模型相对较少，对疾病发病机制的了解有限。目前还没有生物学上相关的系统来筛选有效治疗葡萄膜炎的药物。现有的少数葡萄膜炎小鼠模型没有利用新兴的现代小鼠遗传学资源。我们建议通过产生一组转基因小鼠来创建一种新的葡萄膜炎疾病模型，这些转基因小鼠被设计成在特定的眼组织亚群中表达特定的免疫原性蛋白(来自眼部病原体)。我们预测，当这些病原体攻击时，这些转基因小鼠将发展成眼部炎症性疾病；也就是说，可以通过分子模仿产生合成葡萄膜炎模型。本研究将优化酵母Gal4-UAS反式激活系统用于哺乳动物细胞和转基因小鼠，并利用哺乳动物适应的lac抑制系统增加变阻器功能；创建在眼睛组织特异性启动子控制下表达Gal4转录激活子的转基因小鼠，以及在UAS上游识别序列控制下表达特定蛋白质的转基因小鼠；并分析这些小鼠的后代在病原体和病原体蛋白免疫后的自发性炎症和炎症反应。

DESCRIPTION (provided by applicant): Inflammatory eye disease is a major cause of decreased vision and blindness worldwide. Relatively few animal models of uveitis exist, which has limited understanding of disease pathogenesis. No biologically relevant system for screening of drugs active against uveitis is available. The few existing mouse models of uveitis do not take advantage of the burgeoning set of resources of modem mouse genetics. We propose creating a new model for uveitic disease by generating sets of transgenic mice engineered to express specific immunogenic proteins (derived from ocular pathogens) in specific subsets of ocular tissue. We predict that such transgenic mice will develop ocular inflammatory disease when challenged with those pathogens; that is, a synthetic uveitis model can be generated by molecular mimicry. This research will optimize the yeast Gal4-UAS transactivating system for use in mammalian cells and in transgenic mice, and add rheostat function with the mammalian-adapted lac-repressor system; create transgenic mice expressing the Gal4 transcriptional activator under the control of ocular tissue-specific promoters, and transgenic mice expressing specific proteins under the control of the UAS upstream recognition sequence; and analyze offspring of these mouse strains for spontaneous inflammation as well as inflammation following immunization with pathogen and pathogen protein.