Function and evolution of microRNAs

microRNA的功能和进化

• 批准号: 7123958
• 负责人: Fabio Piano
• 金额: $ 18.68万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-21 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7123958
• 关键词: Nematoda; antisense nucleic acid; binding sites; biochemical evolution; bioinformatics; biotechnology; computational biology; computer program /software; developmental genetics; functional /structural genomics; gene expression; gene induction /repression; helminth genetics; high throughput technology; messenger RNA; microRNAs; molecular biology information system; molecular genetics; nucleic acid inhibitor; oligonucleotides; site directed mutagenesis; species difference

DESCRIPTION (provided by applicant): Recent experiments have shown that the genomes of organisms such as worm, fly, human, and mouse encode hundreds of novel microRNA genes. Many of these small non-coding genes are thought to regulate the translational expression of other genes by binding to partially complementary sites in target messenger RNAs. Initial phenotypic and expression analyses suggest an important role for microRNAs in basic cellular processes such as development, viral response, and apoptosis. Furthermore, many microRNAs are conserved over large evolutionary distances. However, the biological function of most microRNAs is unknown. Although recent computational methods have made progress towards the identification of microRNA targets, no experimental high-throughput method for dissecting microRNA function exists. Our long-term goal is to create a system where bioinformatic and experimental methods are integrated to make possible the high-throughput combinatorial analysis of microRNA function. Here, we propose to setup this system. Specifically, we will (a) employ in silica methods to predict microRNA targets in five different nematode species, (b) use inhibitor oligonucleotides to knock down the function of 20 microRNAs across several nematode species in vivo, and register their loss-of-function phenotypes throughout development, (c) develop a system to validate predicted microRNA targets in vivo and test 25 computationally predicted targets. These data will generate novel insights into the biological function of microRNAs and their potential role in shaping biological diversity. This project is an intensive collaboration between a bioinformatics lab (focused on gene regulatory processes), and an experimental lab (RNAi functional genomics in nematodes)

描述（由申请人提供）：最近的实验表明，生物体（如蠕虫、苍蝇、人类和小鼠）的基因组编码数百种新型microRNA基因。许多这些小的非编码基因被认为通过结合到靶信使RNA中的部分互补位点来调节其他基因的翻译表达。初步的表型和表达分析表明，microRNA在基本的细胞过程，如发育，病毒反应和凋亡中发挥重要作用。此外，许多microRNA在大的进化距离上是保守的。然而，大多数microRNA的生物学功能是未知的。虽然最近的计算方法已经在识别microRNA靶标方面取得了进展，但还没有实验性的高通量方法来解剖microRNA功能。我们的长期目标是建立一个系统，其中生物信息学和实验方法相结合，使高通量microRNA功能的组合分析成为可能。在这里，我们建议建立这个系统。具体而言，我们将（a）采用二氧化硅方法预测五种不同线虫物种中的microRNA靶标，（B）使用抑制剂寡核苷酸在体内敲低几种线虫物种中20种microRNA的功能，并在整个发育过程中记录其功能丧失表型，（c）开发一种系统来验证预测的microRNA靶标，并测试25种计算预测的靶标。这些数据将对microRNA的生物学功能及其在塑造生物多样性中的潜在作用产生新的见解。该项目是生物信息学实验室（专注于基因调控过程）和实验室（线虫RNAi功能基因组学）之间的密切合作