Inducible Transgenic Mouse Model of RNA Toxicity

RNA毒性诱导转基因小鼠模型

• 批准号: 6959625
• 负责人: Mani Subramaniam Mahadevan
• 金额: $ 33.51万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6959625
• 关键词: Lentivirus; RNA splicing; biotechnology; cell differentiation; chloride channels; disease /disorder model; fluorescent in situ hybridization; gene expression; gene mutation; gene therapy; genetically modified animals; immunocytochemistry; laboratory mouse; messenger RNA; myoblasts; myotonic dystrophy; nonhuman therapy evaluation; northern blottings; nucleic acid repetitive sequence; protein kinase; small interfering RNA; tetracyclines; therapy design /development; tissue /cell culture; transfection /expression vector; troponin; western blottings

DESCRIPTION (provided by applicant): Myotonic dystrophy (DM) is the most common inherited neuromuscular disorder in adults. There are two types, DM1 and DM2, both being autosomal dominant disorders caused by expansions of microsatellite repeats within non-coding regions of their respective genes. DM1 is far more common; however both forms of DM are likely to share similar pathogenic mechanisms. The DM1 mutation is an expansion of a CTG triplet repeat in the 3' untranslated region (3'UTR) of the DM protein kinase (DMPK) gene. A prevailing hypothesis in the field is that many aspects of DM are caused by the expression of the mutant mRNA. DM1 and DM2 represent the first examples of toxic RNA mediated disease pathogenesis. We have already developed and characterized extensively, a myoblast cell culture model to clearly demonstrate the toxic effects of the mutant DMPK mRNA on muscle differentiation. To study the hypothesis further, the aims of this proposal are to develop and characterize an inducible transgenic mouse model of RNA toxicity for DM type 1 (DM1) and to develop a siRNA (small interfering RNA) therapeutic approach to get rid of the toxic RNA which can be tested in both our cell culture and transgenic animal models. The development of transgenic mouse models will aid in understanding disease pathogenesis and will also provide a system with which to test out potential therapeutic .strategies. The ability to control gene expression through an inducible system will enable better characterization of and better correlation with the onset and levels of expression of the toxic RNA in DM1 and disease outcomes. Most importantly, because of this property, this model will be one of the first in which we can directly test, at the level of a whole organism, if ablation of expression of the toxic RNA after a period of exposure can reverse its toxic effects. All DM patients have endured exposure to the toxic RNA from birth and thus a model such as this one will be able to provide valuable and relevant insight into this therapeutic strategy.

描述（由申请人提供）：强直性肌营养不良（DM）是成人中最常见的遗传性神经肌肉疾病。有两种类型，DM1 和 DM2，两者都是常染色体显性遗传疾病，由各自基因非编码区域内微卫星重复的扩展引起。 DM1 更为常见；然而，这两种形式的 DM 可能具有相似的致病机制。 DM1 突变是 DM 蛋白激酶 (DMPK) 基因 3' 非翻译区 (3'UTR) 中 CTG 三联体重复的扩展。该领域的一个流行假设是，DM 的许多方面都是由突变 mRNA 的表达引起的。 DM1 和 DM2 代表了有毒 RNA 介导的疾病发病机制的第一个例子。我们已经开发并广泛表征了一种成肌细胞培养模型，以清楚地证明突变型 DMPK mRNA 对肌肉分化的毒性作用。为了进一步研究这一假设，本提案的目的是开发和表征 1 型 DM (DM1) RNA 毒性的诱导转基因小鼠模型，并开发 siRNA（小干扰 RNA）治疗方法来消除有毒 RNA可以在我们的细胞培养物和转基因动物模型中进行测试。 转基因小鼠模型的开发将有助于了解疾病发病机制，并将提供一个测试潜在治疗策略的系统。通过诱导系统控制基因表达的能力将能够更好地表征 DM1 中有毒 RNA 的发作和表达水平以及疾病结果，并与其更好地相关。最重要的是，由于这一特性，该模型将成为我们第一个可以在整个生物体水平上直接测试毒性 RNA 在暴露一段时间后的表达消除是否可以逆转其毒性作用的模型之一。所有 DM 患者从出生起就经历了有毒 RNA 的暴露，因此像这样的模型将能够为这一治疗策略提供有价值且相关的见解。