TRTech-PGR: NSF BSF - Recombining Allelic Diversity via Genome Editing for Dissecting Complex Plant Traits

TRTech-PGR：NSF BSF - 通过基因组编辑重组等位基因多样性以剖析复杂的植物性状

• 批准号: 2034264
• 负责人: Markita Landry
• 金额: $ 60万
• 依托单位: University of California-Berkeley
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2034264&HistoricalAwards=false
• 关键词: TRTech; PGR; NSF; BSF; Recombining

Global sustainability efforts rely on improvements in plant breeding technology particularly for crops of relevance for human consumption such as cereal crops. Wild relatives of domesticated plants harbor many of the genes that enable plants to become robust against stress. However, our ability to identify these genes and allocate their function to plant stress resilience has been difficult due to the low throughput of experiments needed to identify plant resilience genes. A main bottleneck limiting the throughput of testing crop genetic resilience is the ability to deliver biological molecules into plant cells, where the plant cell wall presents a largely impenetrable barrier for the introduction of the molecular biology tools needed for plant genetic mapping. In this project, nanoparticles will be developed to deliver genetic material into plant cells, with a focus on cereal crop barley. These nanoparticles will be chemically functionalized to deliver genome editing cargoes into plants, which will enable probing of specific plant genes in their relevance for plant stress tolerance. The project will seek to identify what genes enable cereal crops to be robust against environmental stress, and to leverage this knowledge to learn about stress tolerance in other crops of relevance for the global food supply. The research above will also support underrepresented undergraduate students as research trainees for the synthesis and characterization of nanomaterials and will develop a student workshop on nanomaterials science. The ability to study genotype to phenotype relationships in plants is critical for identification of plant biotic and abiotic stress genes. Specifically, crop wild relatives harbor genes that confer traits relevant for crop adaptation to climate change. Mapping genotype to phenotype relationships through functional genomics has benefitted greatly from the emergence of CRISPR genome editing technologies. However, the efficiency of genome editing in plants limits the utility of this method for functional genomics. In this project, nanoparticles will be developed for the physiochemical adsorption of CRISPR-based DNA plasmids and, separately, Cas9-gRNA ribonucleoprotein complexes. These nanoparticle-biomolecule cargo complexes will be tested for delivery and genome editing efficacy in model plants and in the cereal crop barley. The ability to deliver CRISPR plasmids in a non-integrating manner or to achieve DNA-free genome editing through Cas9-gRNA complex delivery would enable faster and more precise mapping of plant traits by avoiding transgene integration and the need for transgene segregation. Furthermore, the abiotic nature of nanoparticle-based DNA or protein delivery may enable facile genetic manipulation and functional genomics in other crops of relevance beyond those tested in the current work. In addition to the above efforts, this proposal will support training of undergraduate students from the Society for the Advancement of Chicanos and Native Americans in Science, and the development of a workshop to teach nanomaterials science to students from a broad variety of backgrounds.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.

全球可持续性努力依赖于植物育种技术的改进，特别是与人类消费有关的作物，如谷类作物。驯化植物的野生亲缘植物含有许多使植物能够抵御逆境的基因。然而，我们的能力，以确定这些基因，并分配其功能，以植物胁迫恢复一直是困难的，由于低通量的实验需要确定植物的弹性基因。限制测试作物遗传恢复力的通量的主要瓶颈是将生物分子递送到植物细胞中的能力，其中植物细胞壁对于引入植物遗传作图所需的分子生物学工具呈现出很大程度上不可穿透的屏障。在该项目中，将开发纳米颗粒，将遗传物质输送到植物细胞中，重点是谷类作物大麦。这些纳米颗粒将被化学功能化，以将基因组编辑货物递送到植物中，这将使探测特定植物基因与植物胁迫耐受性的相关性成为可能。该项目将寻求确定哪些基因使谷类作物能够抵御环境压力，并利用这一知识来了解与全球粮食供应相关的其他作物的抗逆能力。上述研究还将支持代表性不足的本科生作为纳米材料合成和表征的研究实习生，并将举办一个关于纳米材料科学的学生讲习班。研究植物基因型与表型的关系对于鉴定植物生物和非生物胁迫基因是至关重要的。具体而言，作物野生近缘种含有赋予作物适应气候变化相关性状的基因。通过功能基因组学绘制基因型与表型的关系极大地受益于CRISPR基因组编辑技术的出现。然而，植物基因组编辑的效率限制了这种方法在功能基因组学中的应用。在这个项目中，纳米颗粒将被开发用于基于CRISPR的DNA质粒和Cas9-gRNA核糖核蛋白复合物的物理化学吸附。这些纳米颗粒-生物分子货物复合物将在模型植物和谷类作物大麦中测试递送和基因组编辑功效。以非整合方式递送CRISPR质粒或通过Cas9-gRNA复合物递送实现无DNA基因组编辑的能力将通过避免转基因整合和对转基因分离的需要来实现植物性状的更快和更精确的作图。此外，基于纳米颗粒的DNA或蛋白质递送的非生物性质可能使其他相关作物中的基因操作和功能基因组学变得容易，而不仅仅是目前工作中测试的那些。除了上述努力外，该提案还将支持对来自促进奇卡诺人和美洲土著人科学进步协会的本科生进行培训，该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查进行评估，被认为值得支持的搜索.

项目成果

The emerging role of nanotechnology in plant genetic engineering

• DOI: 10.1038/s44222-023-00037-5
• 发表时间: 2023-02
• 期刊: Nature Reviews Bioengineering
• 作者: Henry J. Squire;Sophia Tomatz;Elizabeth Voke;Eduardo González-Grandío;M. Landry