Transposon mutagenesis for modeling complex human disease in rats

描述（由申请人提供）：了解复杂人类疾病遗传基础的关键限制之一是良好模型的可用性。实验室大鼠就是这样一种模型，由于其与人类在生理、病理和序列上的相似性而具有巨大的价值。在大鼠的遗传研究中，已经定位了1300多个与疾病相关的数量性状基因座（QTL）。然而，在大鼠研究领域，生理学和遗传学之间存在脱节，因为没有有效的方法来突变和识别基因。我们一直在开发一种新的工具来缩小这一技术差距。我们现在正在建立的初步数据表明，转座子插入诱变使用睡美人（SB）转座子系统可以产生遗传突变的大鼠在比传统方法的效率高50倍的速度。我们相信SB转座的特性，特别是转座子跳跃到染色体内附近序列的倾向（局部跳跃），将使我们能够靶向这些区域进行诱变，并大大加速疾病基因的发现。转座子诱变的另一个强大的应用是癌症基因的发现。在转基因啮齿动物的体细胞中，SB转座子插入突变可以通过罕见但重复的癌基因突变来驱动肿瘤形成。我们可以利用转座子通过研究它们在肿瘤DNA中的插入位点来找到这些癌症基因。我们特别提出并行开发这两种应用以模拟遗传性疾病，通过1）开发在基因组的不同区域中具有转座子的多个转基因菌株以利用QTL区域内的转座子突变的局部跳跃和2）开发用于大鼠体细胞诱变的SB转座子系统。我们选择Dahl盐敏感（SS）近交系大鼠作为我们的模型，因为它是许多研究人员中心血管研究的前沿，我们已经初步将其定性为人类乳腺癌的新模型。我们将联合收割机将SS株和SB转座子突变与几十年的生理和病理特征相结合，建立疾病模型，并发现相关的疾病基因。叙述：研究人员开发诊断测试和治疗复杂人类疾病（如心血管疾病和癌症）的关键是找到相关基因。我们使用我们称之为“遗传模型生物”的东西，如老鼠，来了解不同疾病的病理学，并努力发现与突变有关的基因，或去除它们，并确定发生了什么。我们正在开发一种功能强大的新工具，它在大鼠身上更有效，这样我们就可以加速许多疾病领域的研究，以提高我们所有人的生活质量。

DESCRIPTION (provided by applicant): One of the key limitations to understanding the genetic basis of complex human disease is the availability of good models. The laboratory rat is one such model which has tremendous value because of intensely studied physiological, pathological and sequence similarities to humans. More than 1300 disease-related quantitative trait loci (QTL) have been mapped in genetic studies in the rat. There is a disconnection, however, in the rat research field between physiology and genetics because there are no efficient means to mutate and identify genes. We have been developing a new tool to close this technology gap. We are now building upon preliminary data showing that transposon insertional mutagenesis using the Sleeping Beauty (SB) transposon system can generate heritable mutations in the rat at a rate which is 50-fold more efficient than conventional methods. We believe that properties of SB transposition, specifically the tendency for transposons to jump to nearby sequences within a chromosome (local hopping), will allow us to target these regions for mutagenesis and dramatically accelerate disease gene discovery. Another powerful application of transposon mutagenesis is cancer gene discovery. In somatic cells of transgenic rodents, SB transposon insertional mutagenesis can drive tumor formation by the rare, but repeated mutation of cancer genes. We can use the transposon to find these cancer genes by studying their insertion sites in tumor DNA. We specifically propose to develop these two applications in parallel to model genetic disease by 1) developing multiple transgenic strains which have transposons in different regions of the genome to take advantage of local hopping for transposon mutgenesis within QTL regions and 2) develop the SB transposon system for somatic mutagenesis in the rat. We chose the Dahl salt-sensitive (SS) inbred rat strain as our model because it is at the forefront of cardiovascular research among many researchers and we have preliminarily characterized it as a new model for human breast cancer. We will combine the SS strain and SB transposon mutagenesis with decades of physiological and pathological characterization to create the disease models and discover the disease genes which are involved. Narrative: The key for researchers to develop diagnostic testing and treatments for complex human diseases like cardiovascular disease and cancer is finding the genes that are responsible. We use what we call `genetic model organisms' like rats to understand the pathology of different diseases and work hard to discover genes that are involved by mutating, or removing them, and determining what happens. We are developing a powerful new tool which is much more efficient at doing this in the rat so that we can accelerate research in many disease areas to improve the quality of life for all of us.