NSF Postdoctoral Fellowship in Biology: Methods for Virus-based Plant Genome Editing

NSF 生物学博士后奖学金：基于病毒的植物基因组编辑方法

• 批准号: 2305527
• 负责人: Chandler Meyer
• 金额: $ 24.9万
• 依托单位: Meyer, Chandler Marie
• 依托单位国家: 美国
• 项目类别: Fellowship Award
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2305527&HistoricalAwards=false
• 关键词: NSF; Postdoctoral; Fellowship; Biology; Methods

This action funds an NSF Plant Genome Postdoctoral Research Fellowship in Biology for FY 2023. The fellowship supports a research and training plan in a host laboratory for the Fellow who also presents a plan to broaden participation in biology. The title of the research and training plan for this fellowship to Chandler Meyer is “Methods for Virus-based Plant Genome Editing” The host institution for the fellowship is the University of Minnesota and the sponsoring scientist is Dr. Daniel Voytas.Precision editing of plant genomes has the potential to accelerate crop improvement and advance functional genomics. Considerable progress has been made in this area, however, the most commonly used methods for gene editing in plants often require the use of tissue culture. The drawback of tissue culture is that it requires technical expertise, it is time intensive, and success is limited to a few genotypes. This project aims to develop a viral-based method of genome editing in plants that offers the potential to significantly reduce the need for tissue culture. If successful, the methods developed could be applied to crop species and greatly accelerate the ease of gene editing in crop plants, improving the ability of breeders to create new traits. The results from this work could also have a broad impact on plant biology by streamlining methods for generating mutants for gene function studies. Training objectives include outreach activities to promote community engagement in plant gene editing and plant science, mentoring undergraduate students, and dissemination of plant gene editing techniques by participation in the NSF-funded Plant Genetic Engineering Network (PlantGENE). The most commonly used methods to deliver gene editing reagents to plant cells are Agrobacterium-mediated transformation and biolistic bombardment. To recover a gene-editing plant, tissue culture techniques are needed to regenerate a plant from the edited somatic cells. This is challenging for a few reasons. First, it often requires months of training and development of technical expertise. Second, successful regeneration is often limited to a few genotypes. Lastly, the tissue culture process can result in somaclonal variation. The use of viruses for genome modification in plants offers the potential to ease the tissue culture and reagent delivery bottlenecks. The research objectives of this project are to (1) improve the cargo capacity of TRV-based vectors so that TRV can deliver CRISPR-Cas nucleases to meristematic cells, (2) test the use of miniature CRISPR-Cas nucleases to overcome TRV-based vector cargo capacity limitations, and (3) identify RNA sequences that increase cell-to-cell mobility of sgRNAs and nucleases to promote delivery to the germline. This research will generate knowledge of methods for genomic modification of plants that can be used by plant breeders and researchers to develop novel traits and study gene function. Data generated will be released to the community through submission to public repositories, project web pages, and publications as well as presentations at scientific conferences, seminars, and workshops. Germplasm will be available upon request.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.

这一行动为2023财年NSF植物基因组生物学博士后研究奖学金提供了资金。该奖学金支持在东道主实验室为该研究员制定的研究和培训计划，该研究员还提出了扩大生物学参与度的计划。钱德勒·迈耶的这项研究和培训计划的标题是“基于病毒的植物基因组编辑方法”。该奖学金的主办机构是明尼苏达大学，赞助科学家是丹尼尔·沃伊塔斯博士。植物基因组的精确编辑具有加速作物改良和推进功能基因组学的潜力。在这一领域已经取得了相当大的进展，然而，最常用的植物基因编辑方法通常需要使用组织培养。组织培养的缺点是需要技术专业知识，时间密集，而且成功仅限于少数几种基因。该项目旨在开发一种基于病毒的植物基因组编辑方法，从而有可能显著减少对组织培养的需求。如果成功，开发的方法可以应用于作物物种，并极大地加快作物基因编辑的便利性，提高育种人员创造新特征的能力。这项工作的结果也可能对植物生物学产生广泛的影响，因为它简化了为基因功能研究产生突变的方法。培训目标包括促进社区参与植物基因编辑和植物科学的外联活动，指导本科生，以及通过参加国家科学基金会资助的植物遗传工程网络(PlantGENE)传播植物基因编辑技术。将基因编辑试剂导入植物细胞最常用的方法是农杆菌介导法和生物轰击法。为了恢复基因编辑植物，需要组织培养技术来从编辑后的体细胞再生植物。这是具有挑战性的，原因有几个。首先，它往往需要数月的培训和技术专长的发展。其次，成功的再生往往仅限于少数几个基因类型。最后，组织培养过程可能导致体细胞无性系变异。利用病毒对植物进行基因组修饰，有可能缓解组织培养和试剂输送的瓶颈。本项目的研究目标是(1)提高基于TRV的载体的货物能力，以便TRV能够将CRISPR-Cas核酸酶运送到分生组织细胞；(2)测试使用微型CRISPR-Cas核酸酶来克服基于TRV的载体货物能力的限制；(3)鉴定能够增加sgRNAs和核酸酶的细胞间迁移率的RNA序列，以促进向生殖系的运送。这项研究将产生植物基因组修改方法的知识，植物育种者和研究人员可以使用这些方法来开发新的特征和研究基因功能。产生的数据将通过提交到公共储存库、项目网页和出版物以及在科学会议、研讨会和研讨会上的演讲向社区发布。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。