Genetic Analysis of Conserved microRNAs in C. elegans

描述（由申请人提供）：这项工作的长期目标是确定microRNA如何适应复杂的发育和生理网络。识别由单个microRNA调控的途径和靶点对于了解microRNA在动物发育和人类疾病中的功能至关重要。作为一种遗传上非常温顺的动物，C。线虫为研究microRNA的功能提供了一个理想的模型系统，特别是许多microRNA在蠕虫和人类之间显示出完全或接近完全的保守性。拟议的研究集中在排卵的单一生物过程。排卵是一种钙依赖性节律行为，需要microRNA。这项工作的中心问题是：1）排卵的哪些特定事件需要microRNA？2)生殖细胞或体细胞性腺中有哪些microRNA起着调节排卵的作用？3）这些microRNA调节哪些途径和直接靶点？为了解决这些问题，我们将使用遗传学方法来有条件地降低microRNA的生物合成活性，以确定需要microRNA的特定排卵事件。此外，我们将使用高通量测序来分析在体细胞性腺中表达的microRNA。最后，我们将确定哪些体细胞性腺microRNA有助于控制排卵，并确定它们调节的途径和靶点。这种对单一生物过程的全面分析为确定microRNA在复杂的调控网络中的作用提供了机会。

DESCRIPTION (provided by applicant): The long term goal of this work is to define how microRNAs fit into complex developmental and physiological networks. The identification of pathways and targets regulated by individual microRNAs is vital to understand microRNA function in animal development and in human disease. As an eminently genetically-tractable animal, C. elegans provides an ideal model system in which to study the functions of microRNAs, particularly as many microRNAs show complete or near-complete conservation between worms and humans. The proposed research is focused on the single biological process of ovulation. Ovulation is a calcium-dependent rhythmic behavior that requires microRNAs. The central questions of this work are: 1) what specific events of ovulation require microRNAs? 2) What individual microRNAs function in the germ cells or somatic gonad to regulate ovulation and 3) what pathways and direct targets are regulated by these microRNAs? To address these questions we will use genetic approaches to conditionally reduce microRNA biogenesis activity to determine specific ovulation events that require microRNAs. In addition, we will use high-throughput sequencing to profile microRNAs that are expressed in the somatic gonad. Lastly, we will identify which somatic gonad microRNAs contribute to the control of ovulation and determine the pathways and targets that they regulate. This comprehensive analysis of a single biological process provides an opportunity to define how microRNA's act in complex regulatory networks.