Mammalian Heart and Lung microRNA Deletion and Distribution Resource

描述(由申请人提供)： 大多数对基因功能的生物学分析都集中在编码蛋白质的基因上，这些基因只占人类基因组的1.4%。基因组的其余部分在很大程度上是未被探索的。最近发现了包括microRNAs(MiRNAs)在内的非编码RNA的保守片段，揭示了以前未知的基因组调节层。MiRNAs长度为20-22个核苷酸，在转录后起作用，滴定基因组中至少三分之一的蛋白质编码基因的活性。在某些情况下，miRNAs充当关键通路的开关。在其他情况下，它们起到变阻器的作用，以滴定正常生物中的通路活动，并对外部压力和刺激做出反应。虽然已经发现了450多个人或鼠的miRNAs，但关于这类新的RNAs的大部分知识都来自蠕虫和苍蝇。到目前为止，在小鼠中只有三个miRNAs被报道定向缺失，其中两个在心脏生物学中具有关键功能。这些早期的功能丧失研究揭示了干预人类疾病的新靶点，更多miRNAs的干扰很可能同样具有启发性。 为了促进在这个新兴的和高度重要的生物学领域的发现，我们建议用一种先进的策略来对胚胎干细胞进行基因修饰，以去除-75在进化上保守的心和肺丰富的miRNA基因。这些小鼠将通过一种基于网络的机制向科学界提供，以加速心肺研究几乎每一个方面的进展。这是一项雄心勃勃的努力，但我们相信我们拥有独特的工具和专业知识来实现这一目标。此外，回报可能是巨大的。我们期待着在对基因组这一相对未被探索的部分的理解方面取得重大突破。人类疾病的新小鼠模型可能会揭示人类疾病，在这些疾病中，潜在的编码蛋白质的疾病基因很难在给定的遗传位点内定位。这些疾病包括许多心脏和肺部疾病的发育和产后维持和适应。为了完成这个项目，我们提出了三个具体目标。 具体目的1.制备胚胎干细胞和具有~75个心脏和肺丰富的miRNAs干扰的小鼠。 具体目的2.完成miRNA缺失系的初步鉴定，包括中断验证和内源miRNA表达分析。 具体目标3.向科学界传播关于miRNA干扰的小鼠品系的信息并协调交付。(摘要结束)

DESCRIPTION (provided by applicant): Most biological analyses of gene function have focused on protein-encoding genes, which constitute a mere 1.4% of the human genome. The rest of the genome is largely unexplored. The recent discovery of conserved stretches of non-coding RNAs, including microRNAs (miRNAs), has revealed a previously unrecognized layer of genomic regulation. miRNAs 20-22 nucleotides in length, function post-transcriptionally to titrate the activity of at least one-third of the protein-coding genes in the genome. In some cases, miRNAs function as on-off switches for key pathways. In other situations, they function as a rheostat to titrate the activity of pathways in normal biology and in response to external stresses and stimuli. Although more than 450 human or mouse miRNAs have been identified, most of the knowledge of this novel class of RNAs has come from worms and flies. Targeted deletion in mice has been reported for only three miRNAs to date, with two of them having critical functions in cardiac biology. These early loss-of-function studies have revealed novel targets for intervention in human disease, and it is highly likely that disruption of additional miRNAs will be equally revealing. To catalyze discovery in this emerging and highly significant area of biology, we propose to ablate -75 evolutionarily conserved heart and lung-enriched miRNA genes in the mouse with an advanced strategy to genetically modify embryonic stem cells. These mice will be made available to the scientific community through a web-based mechanism to accelerate advances in virtually every aspect of heart and lung research. This is an ambitious effort, but we believe we have the unique tools and expertise to accomplish it. In addition, the payoff is potentially significant. We expect major breakthroughs in the understanding of this relatively unexplored portion of the genome. New mouse models of human disease could shed light on human diseases in which potential protein-encoding disease genes have been difficult to map within a given genetic locus. These include a host of cardiac and pulmonary diseases of development and post-natal maintenance and adaptation. To accomplish this project, we propose three specific aims. Specific Aim 1. To generate embryonic stem cells and mice with disruption of ~75 heart and lung-enriched miRNAs. Specific Aim 2. To complete an initial characterization of the miRNA deletion lines, including validation of disruption and analysis of endogenous miRNA expression. Specific Aim 3. To disseminate information on and coordinate delivery of miRNA-disrupted mouse lines to the scientific community. (End of Abstract)