MicroRNA Function in the Developing Vertebrate Limb

描述(由申请者提供)：由于对其多样性和在动物物种中流行程度的新发现，人们对小干扰、非编码microRNAs的兴趣激增。尽管它们在这一领域获得了所有的热情，但人们对它们各自的靶基因和生物学功能知之甚少。该方案的目标是结合分子生物学、计算生物信息学和经典胚胎学来确定在脊椎动物肢芽发育中表达的一组microRNAs的功能。特定目标1将测试Let-7E在肢体中发挥作用以抑制不适当的软骨生成的假设。特殊目的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个microRNAs的功能。具体目标3将专注于通过在小鼠中产生Ago2的条件零等位基因并进行微阵列分析来阐明对microRNA相关蛋白ArgAerte 2及其同源microRNAs的需求。这些研究的结果将揭示microRNAs在发育中的胚胎中的功能，并将为可能开发反义RNA技术用于治疗应用的研究开辟道路。

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.