EAGER: Development of an RNA editing tool for probing the function of plant mitochondrial genes and proteins

EAGER：开发用于探测植物线粒体基因和蛋白质功能的 RNA 编辑工具

• 批准号: 1946152
• 负责人: Stephane Bentolila
• 金额: $ 30万
• 依托单位: Cornell University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-11-15 至 2021-10-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1946152&HistoricalAwards=false
• 关键词: EAGER; Development; RNA; editing; tool

This project aims to develop enabling technology for studying essential gene functions in plant mitochondria. In plant cells, like in animals, mitochondria are known as the cellular powerhouses, generating the energy needed for essential activities. Mitochondria contain their own genomes, which are known to be more complex and much larger in plants than in animals; yet, how plant genomes work to produce characteristic RNA and protein products is not well understood. Moreover, genetic tools for studying genes in mitochondria are lacking. Therefore, the tools to be generated by this project could revolutionize the ability to analyze and potentially engineer mitochondrial genes and their products. In addition to this scientific impact, the project will have beneficial educational outcomes by providing undergraduate students with hands-on research training to prepare them to become members of the future STEM workforce.This project aims to resolve the lack of genetic tools to probe mitochondrial gene functions by providing a technology that will allow editing of mitochondrial transcripts at certain nucleotides. The CRISPR revolution in genome editing relies on the use of nucleases, Cas proteins, which act in bacterial immunity by generating double-stranded breaks in invading phage DNAs. Recently, "dead" Cas proteins that have lost their endonuclease activity but retained their binding affinity for nucleic acids have been used to engineer nuclear single base editors acting on either DNA or RNA. This project aims to develop a similar tool for mitochondrial RNA. The impact of this work will provide a novel technology for the field of plant mitochondrial biology. It will open up unprecedented opportunities to understand the function of mitochondrial genes, gene regulatory sequences, and proteins by making precise changes that cannot be made by mutating nuclear genes that encode mitochondrial proteins. This methodology could also provide a way to genetically engineer plant mitochondrial function.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

该项目旨在开发研究植物线粒体重要基因功能的支持技术。在植物细胞中，就像在动物细胞中一样，线粒体被称为细胞动力室，产生基本活动所需的能量。线粒体包含自己的基因组，植物中的线粒体比动物中的更复杂、更大。然而，植物基因组如何产生特征性 RNA 和蛋白质产物尚不清楚。此外，缺乏用于研究线粒体基因的遗传工具。因此，该项目生成的工具可能会彻底改变分析和潜在工程线粒体基因及其产物的能力。除了这种科学影响之外，该项目还将通过为本科生提供实践研究培训来产生有益的教育成果，帮助他们成为未来 STEM 劳动力的成员。该项目旨在通过提供一种允许在某些核苷酸处编辑线粒体转录本的技术来解决缺乏探测线粒体基因功能的遗传工具的问题。基因组编辑领域的 CRISPR 革命依赖于核酸酶、Cas 蛋白的使用，它通过在入侵的噬菌体 DNA 中产生双链断裂来发挥细菌免疫作用。最近，失去核酸内切酶活性但保留核酸结合亲和力的“死”Cas 蛋白已被用来设计作用于 DNA 或 RNA 的核单碱基编辑器。该项目旨在开发一种类似的线粒体 RNA 工具。这项工作的影响将为植物线粒体生物学领域提供一种新技术。它将为理解线粒体基因、基因调控序列和蛋白质的功能提供前所未有的机会，通过实现编码线粒体蛋白质的核基因突变无法实现的精确改变。该方法还可以提供一种对植物线粒体功能进行基因工程的方法。该奖项反映了 NSF 的法定使命，并且通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。