EAGER: Targeted and specific elimination of plant chromosomes

在这个快速变化和干燥的世界中，快速培育和开发适应气候的作物品种是美国国家竞争力和可持续性的必要基础。该提案将CRISPR的基因组编辑从其对一个或几个基因的修饰或改变的优势用途，转变为加速育种迫切需要的基因组规模改变的新工具。这项新技术的发展可以影响多种作物的育种，包括小麦、油菜和花生等异源多倍体作物，这些作物由于基因组的复杂性而具有众所周知的挑战性，但它们是美国主食的关键组成部分。在外联方面，本科生和研究生将在本项目范围内接受先进植物基因组学和分子生物学方面的培训。此外，它将通过面向公众，特别是年轻观众的出版物、演示和/或外展视频传播这项工作的概念、方法和社会影响，为代表性不足的群体接触STEM相关工作提供机会。该战略需要将CRISPR瞄准精心挑选的染色体序列集，以便“免疫”染色体存活，而目标染色体被消除。这导致个体具有特定的、理想的染色体组，例如来自给定父母的染色体组。目的是证明和优化加入特定的染色体消除和工程消除的亲本染色体组在番茄，有效地导致单倍体诱导。使用拟南芥和番茄这两种模型，这项工作涉及开发用于鉴定登录和染色体特异性CRISPR靶点的方法，用于快速验证这些靶点，以及用于基因型和核型特异性驱动的工程化。通过定向消除染色体（TEC）对作物基因组进行基因组规模的改变，有可能使单倍体诱导、异源多倍体基因组操作以及在杂交后代中选择特定的全基因组单倍型构型的广泛应用成为可能。该奖项反映了NSF的法定使命，并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。

In this rapidly changing and drying world, the rapid breeding and development of climate-adapted crop varieties is a necessary foundation of US national competitiveness and sustainability. This proposal takes genome editing by CRISPR from its preponderant use of modification or alteration of one or few genes, to a novel tool for genome-scale alterations that are urgently needed to accelerate breeding. The development of this novel technology can impact the breeding of multiple crops, including allopolyploid ones, such as wheat, canola, and peanut, that can be notoriously challenging due to their genomic complexity, but are critical components of US’s staple foods. In terms of outreach, undergraduate and graduate students will be trained in advanced plant genomics and molecular biology in the context of this project. Additionally, it will provide opportunities for exposing underrepresented groups to STEM-related work through dissemination of the concepts, methods and societal implications of this work through publications, presentations and / or outreach videos geared towards the general public and, specifically, younger audiences.The strategy entails targeting CRISPR to hand-picked sets of chromosomal sequences, so that “immune” chromosomes survive, while targeted chromosomes are eliminated. This results in individuals with specific, desirable chromosome sets, such as those from a given parent. The objectives are to demonstrate and optimize accession-specific chromosome elimination and to engineer elimination of a parental chromosome set in tomato, effectively resulting in haploid induction. Using both the model arabidopsis and the crop tomato, the work involves development of methods for identification of accession- and chromosome-specific CRISPR targets, for rapid validation of these targets, and for engineering of genotype- and karyotype-specific drives. Genome-scale alterations of crop genomes through Targeted Elimination of Chromosomes (TEC), could potentially enable broad application of haploid induction, manipulation of allopolyploid genome, and selection of specific genome-wide haplotypic configurations in hybrid progeny.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.