Genome-wide expression profiling of miRNAs with single cell resolution

描述（由申请人提供）：MicroRNA是基因调控因子，其丰度（约占动物基因组中基因的1%）最近才变得明显。 尽管它们丰富，但对其表达模式和它们发挥作用的生理背景知之甚少。 迄今为止，已经进行了功能研究的少数动物miRNAs表明，miRNAs嵌入在决定细胞命运规范的基因调控级联中。 此外，可以想象，miRNA可以遵循转录因子的先例并以组合方式起作用，使得在给定细胞类型中表达的miRNA的特定组合可以限定并主动确定细胞类型的身份。 为了证实这一假设，研究者将使用线虫C. elegans作为一个模型系统。 他的研究将（1）建立一个miRNA表达谱的目录，（2）为未来miRNA敲除的表型分析提供指导，（3）为“miRNA密码子”的概念提供支持性证据，从而提供关于miRNA在发育生物体中产生细胞多样性的功能的进一步可验证的假设。 此外，它将（4）验证“传感器策略”，以确定miRNA表达谱在C。elegans，以及（5）构建潜在有用的GFP细胞命运标记的工具包。

DESCRIPTION (provided by applicant): MicroRNAs are gene regulatory factors whose abundance (~1% of genes in animal genomes) has only recently become apparent. In spite of their abundance, little is known about their expression patterns and the physiological contexts in which they function. The few animal miRNAs that have been functionally studied to date suggest that miRNAs are embedded in gene regulatory cascades that determine cell fate specification. Moreover, it is conceivable that miRNAs may follow the precedent of transcription factors and act in a combinatorial manner, such that specific combinations of miRNAs expressed in a given cell type may define and actively determine the identity of the cell type. To substantiate this hypothesis, the investigator will determine miRNA expression profiles both on a genome-wide level as well as with a single cell resolution using the nematode C. elegans as a model system. His studies will (1) build a catalogue of miRNA expression profiles, (2) provide a guideline for future phenotypic analysis of miRNA knockouts, and (3) provide supportive evidence for the concept of "miRNA codes", and therefore supply further testable hypothesis about miRNA function in the generation of cellular diversity in a developing organism. Moreover, it will (4) validate the "sensor strategy" to determine miRNA expression profiles in C. elegans, and (5) build a toolkit of potentially useful gfp cell fate markers.