EAGER: Development of the PARE approach for genome-wide profiling of mRNA decay in Arabidopsis

EAGER：开发用于拟南芥全基因组 mRNA 衰减分析的 PARE 方法

• 批准号: 0946326
• 负责人: Pamela Green
• 金额: $ 29.97万
• 依托单位: University of Delaware
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0946326&HistoricalAwards=false
• 关键词: EAGER; Development; PARE; approach; genome

The regulation of genes can occur at many steps, some of which are routinely investigated thousands or tens of thousands at a time using genomic approaches. Many of these methods focus on mRNA because intact mRNA is essential for gene function. This project will expand the capabilities of PARE (Parallel Analysis of RNA Ends), a new genomics method that allows gene regulation to be studied by examining pieces of mRNA molecules that cells are throwing away. Sifting through this cellular trash, millions of molecules at a time, will be a powerful way to identify new types of regulation that were missed previously. PARE was previously very successful for detecting mRNAs whose destruction was caused by regulators called miRNAs. This work will develop PARE for detection of a much broader range of mRNA pieces using the reference plant Arabidopsis. The patterns of mRNA pieces produced in different tissues and when plants are exposed to adverse environmental conditions will also be investigated. This should result in the identification of genes that are controlled at the level of mRNA destruction by drought, high temperature and more. The work should represent a major advance because the cellular trash from mRNA destruction has been very difficult to study previously and PARE is broadly applicable as discussed below.Broader impacts of the proposed researchThe development of PARE as a next-generation technology for the study of gene regulation will be far-reaching in many ways. The use of PARE to the study environmental stress responses, may suggest new ways to make plants stress-resistant. It will also help interpret existing data from other approaches that can not separate intact mRNAs from mRNA pieces. More importantly, because mRNA and its destruction are critical in all organisms from bacteria to man, this project will make it possible to use PARE in all of these systems to study how mRNA destruction is regulated under any condition. The data will be made publicly available worldwide on our user-friendly websites with tools for its analysis. A postdoctoral associate will be mentored under the project, and it should also lead to new collaborations and expose other scientists and potentially the general public to the work.

基因的调控可以发生在许多步骤中，其中一些步骤通常使用基因组方法一次研究数千或数万个步骤。其中许多方法都集中于 mRNA，因为完整的 mRNA 对于基因功能至关重要。该项目将扩展 PARE（RNA 末端并行分析）的功能，这是一种新的基因组学方法，可以通过检查细胞丢弃的 mRNA 分子片段来研究基因调控。一次筛选出数百万个分子的细胞垃圾，将是识别以前遗漏的新型调控的有效方法。 PARE 以前在检测 mRNA 方面非常成功，这些 mRNA 的破坏是由称为 miRNA 的调节因子引起的。这项工作将开发 PARE，以使用参考植物拟南芥来检测更广泛的 mRNA 片段。还将研究不同组织中以及植物暴露于不利环境条件时产生的 mRNA 片段的模式。这应该能够鉴定出在干旱、高温等因素导致的 mRNA 破坏水平上受到控制的基因。这项工作应该代表着一项重大进展，因为以前很难研究 mRNA 破坏产生的细胞垃圾，而 PARE 具有广泛的适用性，如下所述。拟议研究的更广泛影响 PARE 作为基因调控研究的下一代技术的发展将在许多方面产生深远的影响。使用 PARE 来研究环境胁迫反应，可能会提出使植物抵抗胁迫的新方法。 它还将有助于解释无法将完整 mRNA 与 mRNA 片段分开的其他方法的现有数据。 更重要的是，由于 mRNA 及其破坏对于从细菌到人类的所有生物体都至关重要，因此该项目将使在所有这些系统中使用 PARE 来研究在任何条件下如何调节 mRNA 破坏成为可能。这些数据将在我们用户友好的网站上向全球公开，并提供分析工具。一名博士后助理将在该项目下接受指导，它还应该带来新的合作，并使其他科学家和潜在的公众接触到这项工作。