Tools for the Analysis of MicroRNA Function

描述（由申请人提供）：称为微小RNA（miRNA）的RNA分子可以影响靶基因的表达。据认为，哺乳动物miRNA改变其靶蛋白的表达，同时使同源mRNA的表达保持不变。 这些非蛋白质编码的miRNA不仅表达不同，而且直接参与重要的生物学过程，如发育和B细胞分化。预测存在200-255个哺乳动物miRNA基因（对应于所有蛋白质编码基因的1%的数量）。有趣的是，超过52%的miRNA基因位于染色体脆性位点，其中一些与疾病有关。由于每个miRNA都有可能在几个细胞靶点上发挥作用，因此它们的调控复杂性可能是巨大的。 虽然许多预测的miRNA已经被克隆，但预测的miRNA靶点尚未被评估，并且大多数哺乳动物miRNA目前还没有确定的生物学作用。 我们建议开发的工具，将有助于识别的miRNA功能和确认/识别内源性miRNA的目标。我们计划建立一个miRNA报告系统、miRNA抑制剂文库、miRNA表达载体文库或与已知人类miRNA相对应的化学合成miRNA文库。 利用这些试剂，我们将开始分析miRNA调控的基因通路，并鉴定参与各种细胞过程的miRNA，包括凋亡、细胞周期、血管生成、增殖、分化和DNA修复。结合所得数据将提供miRNA如何影响人类细胞的全局视图，并可能揭示潜在的治疗靶点。

DESCRIPTION (provided by applicant): RNA molecules known as microRNA (miRNA) can influence the expression of target genes. It is thought that mammalian miRNA change the expression of their targets protein while leaving the expression of the cognate mRNA unmodified. These non-protein-encoding miRNA are not only differently expressed but are directly involved in important biological processes such as development and B-cell differentiation. Between 200-255 mammalian miRNA genes are predicted to exist (a number that corresponds to 1 % of all protein encoding genes). Interestingly over 52% of miRNA genes are located in chromosomal fragile sitessome of which are associated with disease. Since each miRNA has the potential to function on several cellular targets their regulatory complexity is potentially enormous. While many of the predicted miRNA have been cloned, the predicted miRNA targets have not yet been evaluated and most mammalian miRNA currently have no identified biological role. We propose to develop the tools that will facilitate the identification of miRNA function and confirm/identify endogenous miRNA targets. We propose to develop a miRNA reporter system, library of miRNA inhibitors and library of miRNA expression vectors or chemically synthesized miRNA corresponding to the known human miRNA. Using these reagents, we will begin to analyze miRNA regulated gene pathways and identify miRNA that participate in a variety of cellular processes, including apoptosis, cell cycle, angiogenesis, proliferation, differentiation, and DNA repair. Combining the resulting data will provide a global view of how miRNA influence human cells and might reveal potential therapeutic targets.