Methods of systematic microRNA target validation and identification

系统性 microRNA 靶标验证和识别方法

• 批准号: 8516466
• 负责人: YIN-YUAN MO
• 金额: $ 17.5万
• 依托单位: UNIVERSITY OF MISSISSIPPI MED CTR
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8516466
• 关键词: Algorithms; Biogenesis; Cell physiology; Collection; Complementary DNA; Computer Assisted; Databases; Disease; Functional RNA; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Human; Lead; Learning; Libraries; Malignant Neoplasms; Mediating; Messenger RNA; Methods; MicroRNAs; Molecular; Neck; Normal Cell; Pathway interactions; Phenotype; Plasmids; Play; Process; Regulation; Regulator Genes; Repression; Research; Role; Science; Seeds; Sequence Homology; Source; Specificity; System; Technology; Testing; Translational Repression; Untranslated Regions; Validation; Work; base; human disease; innovation; mRNA Transcript Degradation; novel; research study; success; tool

DESCRIPTION (provided by applicant): It is well known that microRNAs could play a fundamental role in regulation of diverse cellular functions. As key gene regulators, microRNAs work through a posttranscriptional repression mechanism. Increasing evidence indicates that deregulation of microRNA expression could lead to a variety of disorders including human cancer. Although significant progress has been made in the past years in discovery of microRNAs and their biogenesis, and their role in many cellular phenotypes, it is not fully understood how microRNAs exert their cellular functions because a single microRNA can have hundreds of targets. Hence, identification of microRNA targets is a critical step toward understanding of molecular mechanisms of microRNA-mediated gene expression in normal and disease processes. Currently, this largely relies on computer-aided algorithms, which unfortunately are still unable to provide a precise picture of microRNA regulatory networks, and thus the predicted targets need further experimental validations. It is evident that target validation is a bottle neck in our effort to dissect microRNA pathways. In this application, we propose to develop a novel selection method for microRNA target validation and identification through two complementary approaches. The first approach is to determine microRNA/mRNA interactions using our pre-microRNA collection against a specific target cloned in our selection plasmid; the second approach is to determine microRNA/mRNA interactions using a 3'-UTR (untranslated region) library against a specific microRNA. We believe that our selection method is innovative, simple and powerful. An additional benefit of this method will allow us to determine whether there are any new features, besides the seed sequence homology, which could contribute to the specificity of microRNA targeting. Accordingly, this study will greatly enhance our understanding of microRNA targeting and gene regulation by providing a valuable research tool.

描述（由申请人提供）：众所周知，microRNA可以在调节各种细胞功能的调节中起基本作用。作为关键基因调节剂，microRNA通过转录后的抑制机制起作用。越来越多的证据表明，microRNA表达的放松管制可能导致包括人类癌症在内的多种疾病。尽管过去几年在发现microRNA及其生物发生及其在许多细胞表型中的作用方面取得了重大进展，但由于单个microRNA可以具有数百个靶标，因此尚未完全了解microRNA如何发挥其细胞功能。因此，鉴定microRNA靶标是了解正常和疾病过程中microRNA介导的基因表达的分子机制的关键步骤。当前，这主要依赖于计算机辅助算法，不幸的是，该算法仍然无法提供MicroRNA调节网络的精确图片，因此预测的目标需要进一步的实验验证。显然，目标验证是我们努力剖析microRNA途径的瓶颈。在此应用程序中，我们建议通过两种互补方法开发一种新颖的选择方法，以用于microRNA目标验证和识别。第一种方法是使用我们的微米收集前收集来确定MicroRNA/mRNA相互作用，以针对我们选择质粒中克隆的特定靶标。第二种方法是使用针对特定microRNA的3'-UTR（未翻译区域）文库来确定microRNA/mRNA相互作用。我们认为，我们的选择方法是创新，简单且强大的。这种方法的另一个好处将使我们能够确定除种子序列同源性外是否有任何新功能，这可能有助于microRNA靶向的特异性。因此，这项研究将通过提供宝贵的研究工具来大大增强我们对MicroRNA靶向和基因调节的理解。