MicroRNA Function in the Developing Vertebrate Limb

描述（由申请人提供）：由于对小干扰、非编码microrna的多样性和在动物物种中的普遍存在的新发现，对它们的兴趣已经爆发。尽管它们在该领域获得了极大的热情，但人们对它们的个体靶基因和生物学功能知之甚少。本研究的目的是通过结合分子生物学、计算生物信息学和经典胚胎学来确定在发育中的脊椎动物肢体芽中表达的一组microrna的功能。Specific Aim 1将测试let-7e在肢体中抑制不适当软骨形成的假设。Specific Aim 2将使用类似的实验方法来研究在脊椎动物肢体中强烈表达的另外五种microrna的功能。特异性目标3将通过在小鼠中产生Ago2的条件空等位基因并进行微阵列分析，重点阐明对microRNA相关蛋白Argonaute 2及其同源microRNA的需求。这些研究结果将揭示microrna在发育中的胚胎中的功能，并将开辟研究途径，可能允许发展用于治疗应用的反义RNA技术。

DESCRIPTION (provided by applicant): The interest in small interfering, non-coding microRNAs has exploded due to emerging discoveries of their diversity and prevalence across animal species. For all of the enthusiasm that they have garnered in the field, very little is known of their individual target genes and biological functions. The goal of this proposal is to determine the function of a set of microRNAs expressed in the developing vertebrate limb bud by combining molecular biology, computational bioinformatics, and classical embryology. Specific Aim 1 will test the hypothesis that let-7e functions in the limb to inhibit inappropriate chondrogenesis. Specific Aim 2 will use similar experimental methods to investigate the functions of five additional microRNAs expressed strongly in the vertebrate limb. Specific Aim 3 will focus on elucidating the requirement for the microRNA assocated protein Argonaute 2 and its cognate microRNAs by generating a conditional null allele of Ago2 in the mouse and performing a microarray analysis. The results of these investigations will reveal functions of microRNAs in the developing embryo and will open avenues to research that may allow development of antisense RNA technology for therapeutic applications.