Functional Analysis of microRNAs in C. elegans spermatogenesis

7.项目摘要 小RNA，包括microRNA，已被证明在调节细胞凋亡中起关键作用。 生殖细胞系中的基因表达。然而，个体调节的特定途径和靶点 精子发生过程中的microRNA在很大程度上仍然未知。直接识别 由单个microRNA调控的靶标仍然是该领域的核心挑战。在这份协议中，我们 建议确定生殖系microRNAs的直接靶点，这些microRNAs在调节 精子发生我们使用C.优雅的，这一直是工具，在定义 个体microRNA调控发育途径中靶点的核心原则。于过往 通过这项工作，我们和其他人已经确定了在C.线虫生殖细胞， 体细胞性腺细胞此外，我们已经确定了一组微RNA，其功能是调节精子 阵然而，它们调节的途径和靶点在很大程度上仍然没有特征。这 工作将解决两个中心问题：1）与生殖系相关的靶mRNA是什么？ 特定的Argonaute蛋白？2)microRNAs是如何通过靶mRNA的3'非翻译区进行调控的 控制精子生成的作用这项工作将为今后的研究提供基础， 全面定义在生殖系中起作用的microRNA靶点，以调节生殖细胞的发育过程。 精子发生

7. PROJECT SUMMARY Small RNAs, including microRNAs, have been shown to play critical roles in the regulation of gene expression in the germ line. However, the specific pathways and targets regulated by individual microRNAs in the process of spermatogenesis remain largely unknown. The identification of direct targets regulated by individual microRNAs remains a central challenge in the field. In this grant, we propose to identify direct targets of germline microRNAs that function in the regulation of spermatogenesis. We are using the model system of C. elegans, which has been instrumental in defining the core principles of individual microRNA regulation of targets in developmental pathways. In prior work, we and others have identified the set of microRNAs that are expressed in C. elegans germ cells and somatic gonad cells. Additionally, we have identified a set of microRNAs that function to regulate sperm formation. However, the pathways and targets that they regulate remain largely uncharacterized. This work will address two central questions: 1) What are the target mRNAs that associate with a germline specific Argonaute protein? 2) How does regulation by microRNAs through the 3’ UTR of target mRNAs act to control spermatogenesis? This work will provide a foundation for future investigation to comprehensively define the microRNA targets that act in the germ line to regulate the process of spermatogenesis.