TRANSFORM-PGR: Mining the compact Utricularia genome as source of novel regulatory elements for crop biotechnology

TRANSFORM-PGR：挖掘紧凑的狸藻基因组作为作物生物技术新型调控元件的来源

• 批准号: 1759837
• 负责人: Wayne Parrott
• 金额: $ 48.78万
• 依托单位: University of Georgia Research Foundation Inc
• 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: 2019-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1759837&HistoricalAwards=false
• 关键词: TRANSFORM; PGR; Mining; compact; Utricularia

The goal of this project is to advance plant genetic engineering technology and enable next-generation research and plant-improvement to help meet the upcoming challenges in agriculture. The need for increased but sustainable food production needs to be addressed through many approaches. Bioengineering can help address human food security through crop improvement. These improved crops will range from varieties highly adapted to diverse climates to crops with improved nutrition, that are better able to resist pests, use less water and fertilizer, or make the raw materials for bioplastics and other biodegradable materials. To make these plants a reality, it is first necessary to develop the ability to bioengineer multiple traits into plants. Current technology can place many genes close together and insert them into a crop; however, placing these genes close to each other causes them to interfere with each other. Hence, plants typically put a lot of space between their genes, but there is at least one plant known, the bladderwort, that has solved the problem of gene spacing its genome has many genes that are close together, but do not interfere with one another. The bladderwort also contains shortened versions of other DNA sequences that control when and where genes are expressed. This project uses bioinformatics to identify the bladderwort DNA sequences that allow for close spacing of genes, and to isolate the shorter versions of DNA control sequences. The function of each sequence will be validated empirically, and then made publicly available. The research team will also use the bladderwort sequences to find similar control sequences in other plants, including important crop species.Currently, the lack of appropriate DNA regulatory elements can make it difficult to get coordinated expression of multiple genes cloned into one vector. A few DNA insulators have been identified in plants that prevent interference from neighboring genes, but these insulators are so large their use makes vectors impossibly large. The compact genome (just 82 Mb) of Utricularia gibba (bladderwort) has solved many of these expression issues as it evolved. Expression patterns makes it clear this species has compact promoters, terminators, and insulators, as well as bidirectional promoters. Identification, cloning and validation of these elements and their function will provide a set of tools that will enable the routine use of multiple gene engineering. The DNA elements identified in this project will be made publicly available via AddGene, and thus contribute to a new generation of genetic engineering vectors. The researchers will also use the bladderwort sequences to identify similar conserved regulatory elements across angiosperms.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.

该项目的目标是推进植物基因工程技术，使下一代研究和植物改良能够帮助应对即将到来的农业挑战。需要通过许多办法解决增加但可持续的粮食生产的需要。生物工程可以通过作物改良来帮助解决人类粮食安全问题。这些改良作物将从高度适应不同气候的品种到营养改善的作物，这些作物能够更好地抵抗虫害，使用更少的水和肥料，或者制造生物塑料和其他可生物降解材料的原材料。要使这些植物成为现实，首先必须发展将多种性状生物工程改造成植物的能力。目前的技术可以将许多基因放在一起，并将它们插入作物中；然而，将这些基因放在一起会导致它们相互干扰。因此，植物通常会在它们的基因之间留出很大的间隔，但至少有一种已知的植物--水仙--解决了基因间隔的问题。它的基因组有许多相互接近但不相互干扰的基因。水仙还含有其他DNA序列的缩短版本，这些DNA序列控制基因表达的时间和地点。该项目使用生物信息学来识别能够实现近距离基因间隔的水仙DNA序列，并分离出较短版本的DNA控制序列。每个序列的功能都将经过经验验证，然后公开可用。研究小组还将利用水仙序列在其他植物中寻找类似的控制序列，包括重要的作物物种。目前，缺乏适当的DNA调控元件可能会使克隆到一个载体中的多个基因难以协调表达。已经在植物中发现了一些DNA绝缘体，可以防止邻近基因的干扰，但这些绝缘体太大了，以至于使用这些绝缘体使载体变得不可能大。短小的基因组(只有82Mb)在进化过程中解决了许多这样的表达问题。表达模式表明，该物种有紧密的启动子、终止子和绝缘子，以及双向启动子。鉴定、克隆和验证这些元件及其功能将提供一套工具，使多基因工程的常规使用成为可能。该项目中确定的DNA元素将通过Addgene公开提供，从而为新一代基因工程载体做出贡献。研究人员还将使用水仙序列来识别被子植物中类似的保守调控元件。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。