Collaborative Research: TRTech-PGR TRACK: Discovery and characterization of small CRISPR systems for virus-based delivery of heritable editing in plants.

合作研究：TRTech-PGR TRACK：小型 CRISPR 系统的发现和表征，用于基于病毒的植物遗传编辑传递。

• 批准号: 2334027
• 负责人: Steven Jacobsen
• 金额: $ 120万
• 依托单位: University of California-Los Angeles
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-02-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2334027&HistoricalAwards=false
• 关键词: Collaborative; Research; TRTech; PGR; TRACK

Given current crop productivity projections, agricultural practices will be inadequate to meet future global food demands. Humanity’s ability to address this problem will largely depend on the efficiency with which novel genetic diversity can be created and introduced into plant breeding programs. New genome editing techniques allow for a more precise introduction of genetic diversity, but the current methods are very slow and inefficient and only work in some crop species. This project aims to discover novel genome editing systems that can be more widely and quickly deployed in a large variety of crop species, to enhance plant breeding to make crops with higher yields and resistance to drought, pests, and extreme temperatures. The new tools will also be useful for orphan crop species that do not receive sufficient attention from the plant biotechnology industry. The project will also include a structured approach to involving undergraduate student researchers from diverse backgrounds through different programs at the two participating University of California campuses, UCLA and UC Berkeley. The project will also enable postdocs and graduate students on the project to gain experience in training students from diverse backgrounds.Recent progress in genome editing technology is poised to accelerate plant breeding programs by allowing for the precise introduction of specific changes to important plant genes. Despite this advance, a primary bottleneck remains: fast and effective delivery of the gene editing reagents into crop plants. The most common methods of delivery are to encode RNA-guided genome editors (e.g. CRISPR-Cas enzymes) within transgenes and use tissue culture and transformation approaches or to introduce CRISPR protein and guide RNAs directly into plant cells followed by tissue culture to regenerate plants. However, tissue culture methods require considerable time, resources, and technical expertise, and can cause unintended changes to the genome and epigenome. Furthermore, regenerating plants from tissue culture only works in a limited number of plant species and genotypes. Plant viruses are ideal vectors for the delivery of CRISPR systems to whole plants without the use of plant transformation or tissue culture. However, most viruses have a very small cargo capacity, which is insufficient to accommodate currently used CRISPR systems. We propose to discover and characterize hypercompact CRISPR systems that are small enough to be encoded in plant viruses for easy and fast editing of whole plants, which could be used in a wide range of important crop species. We will screen a large number of candidate compact gene editing systems from metagenomics data and test these systems in bacterial cells and plant cells. The goal will be to find hypercompact editing systems that match or exceed the efficiency of current large systems.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.

鉴于当前的作物生产力项目，农业实践将不足以满足未来的全球粮食需求。人类解决这个问题的能力在很大程度上取决于可以创建新的遗传多样性并引入植物育种计划的效率。新的基因组编辑技术可以更精确地引入遗传多样性，但是当前的方法非常缓慢且效率低下，并且在某些作物物种中仅起作用。项目旨在发现新型的基因组编辑系统，这些系统可以更广泛，更快地部署在各种农作物物种中，以增强植物繁殖，从而使农作物产量更高，对干旱，害虫和极端温度具有更高的产量和抗性。新工具也将对植物生物技术行业没有足够关注的孤儿作物物种有用。该项目还将包括一种结构化方法，通过在加州大学加州大学洛杉矶分校和加州大学哥伦比亚分校的两个参与大学校园的不同计划中参与来自不同背景的本科生研究人员。该项目还将使该项目的博士后和研究生能够获得培训来自潜水员背景的学生的经验。基因组编辑技术的进展被毒死以通过允许对重要植物基因的特定更改进行精确引入特定的变化来加速植物育种计划。尽管有这一进步，但仍存在主要的瓶颈：将基因编辑试剂快速有效地递送到作物植物中。最常见的传递方法是在翻译中编码RNA引导的基因组编辑器（例如CRISPR-CAS酶），并使用组织培养和转化方法或引入CRISPR蛋白，并将RNA直接引导到植物细胞中，然后是组织培养物以再生植物。但是，组织培养方法需要考虑时间，资源和技术专长，并可能对基因组和表观基因组引起意外变化。此外，从组织培养物中再生植物只能在有限数量的植物物种和基因型中起作用。植物病毒是将CRISPR系统传递给整个植物的理想媒介，而无需使用植物转化或组织培养。但是，大多数病毒的货物容量很小，这不足以容纳当前使用的CRISPR系统。我们建议发现和表征超大型CRISPR系统，这些系统足够小，可以在植物病毒中编码，以便于整个植物的易于编辑，这些植物可以在各种重要的农作物物种中使用。我们将从宏基因组学数据中筛选大量候选紧凑基因编辑系统，并在细菌细胞和植物细胞中测试这些系统。目的是找到匹配或超过当前大型系统效率的超紧缩编辑系统。该奖项反映了NSF的法定任务，并使用基金会的知识分子优点和更广泛的影响审查标准来诚实地通过评估来诚实地支持。