Expanding the range of barley and potato gene editing tools.

扩大大麦和马铃薯基因编辑工具的范围。

• 批准号: 2891492
• 金额: --
• 依托单位: University of Dundee
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2891492
• 关键词: Expanding; range; barley; potato; gene

Generating new high-yielding, safe, nutritious, climate-resilient crops is essential to alleviate the many well-documented pressures on agriculture. Gene-editing (GE) or precision breeding is a remarkable tool that is fundamentally different from established conventional breeding and GM methods. Appropriate genetic targets that could potentially demonstrate social, environmental and economic value remain scarce in crop species. The student will explore targets for GE in barley and potato, providing the appropriate background and rationale to explain why these targets could be important. The output will be an inventory of genes that could be altered using precision breeding to improve the performance of barley and potato. Selected key gene targets will be used for practical exemplar GE projects. We have had some success in generating barley edited lines using the established CRISPR/Cas9 system. New exemplar gene targets may be created using our established methods. However, GE is an emerging and rapidly evolving technology and the possibility to generate any base substitution using base editing or by prime editing expands plant gene editing opportunities further. Cytosine and adenine base editors allow the conversion of C/G to T/A and A/T to G/C substitutions. The prime editing system involves the use of a Cas9 nickase-reverse transcriptase (M-MLV RT) fusion proteins paired with a pegRNA composed of a gRNA, a primer binding site (PBS) and a reverse transcriptase template1,2,3. The studentship will allow the establishment and development of these new state of the art editing procedures in our crops. In plants, there can be an intermediary step that creates a transgenic plant to contain the CRISPR-Cas9 components to allow the mutation to occur. There are methods that remove this intermediary step altogether by using transitory delivery of the assembled Cas9 protein-gRNA into the cell to generate the mutation, usually through delivery to plant protoplasts. This has great advantages in editing crop elite lines directly without the need for transformation. But protoplasts represent great challenges too in isolating protoplasts from the most suitable tissues and regeneration of plants from protoplasts. The student will have the opportunity to develop consistent protoplast-based methods of delivery within our crop species of interest.Application of GE remains controversial in Europe and subject to debate at all levels, from the public through to the scientific community and policy makers. This studentship is an ideal way to engage in this debate and explore the social, environmental and nutritional benefits of GE. The studentship will provide training in GE methodology in Hutton's established Biotechnology facility with staff experienced in plant transformation. The supervisors have good connections with other teams working in this area with possible opportunities for relevant training at other institutions. The PhD project will provide the student with multiple basic skills in molecular biology, microscopy, plant cell biology and knowledge exchange. The student will have the advantage of taking part in multiple training opportunities through Eastbio, Dundee University and Hutton. For example, at Hutton, there will opportunities to take part in advanced statistical analysis and R training.

培育新的高产、安全、营养、气候适应型作物对于缓解许多有据可查的农业压力至关重要。基因编辑（GE）或精准育种是一种非凡的工具，与现有的传统育种和转基因方法有着根本的不同。在作物物种中，能够潜在展示社会、环境和经济价值的适当遗传目标仍然稀缺。学生将探索大麦和马铃薯的基因工程目标，提供适当的背景和理由来解释为什么这些目标可能很重要。产出将是一份基因清单，可以通过精准育种来改变这些基因，以提高大麦和马铃薯的性能。选定的关键基因目标将用于实际的 GE 示例项目。我们在使用已建立的 CRISPR/Cas9 系统生成大麦编辑品系方面取得了一些成功。可以使用我们已建立的方法创建新的示例性基因靶标。然而，GE 是一项新兴且快速发展的技术，使用碱基编辑或引物编辑产生任何碱基替换的可能性进一步扩大了植物基因编辑的机会。胞嘧啶和腺嘌呤碱基编辑器允许将 C/G 转换为 T/A 以及将 A/T 转换为 G/C 替换。引物编辑系统涉及使用 Cas9 切口酶-逆转录酶 (M-MLV RT) 融合蛋白与由 gRNA、引物结合位点 (PBS) 和逆转录酶模板组成的 pegRNA 配对1,2,3。学生奖学金将使我们能够在农作物中建立和开发这些最先进的新编辑程序。在植物中，可能有一个中间步骤来创建含有 CRISPR-Cas9 组件的转基因植物，以允许突变发生。有一些方法可以完全消除这一中间步骤，即通过将组装好的 Cas9 蛋白-gRNA 暂时递送到细胞中以产生突变（通常是通过递送到植物原生质体）。这对于直接编辑作物优良品系而不需要进行转化具有很大的优势。但原生质体在从最合适的组织中分离原生质体以及从原生质体再生植物方面也面临着巨大的挑战。学生将有机会在我们感兴趣的作物物种中开发一致的基于原生质体的递送方法。GE的应用在欧洲仍然存在争议，并且受到从公众到科学界和政策制定者的各个层面的争论。该奖学金是参与这场辩论并探索 GE 的社会、环境和营养效益的理想方式。该奖学金将在 Hutton 已建立的生物技术设施中提供 GE 方法学培训，并配备在植物转化方面经验丰富的工作人员。主管与该领域的其他团队有良好的联系，并有可能在其他机构接受相关培训的机会。博士项目将为学生提供分子生物学、显微镜学、植物细胞生物学和知识交流方面的多种基本技能。学生将有机会通过 Eastbio、邓迪大学和赫顿参加多种培训机会。例如，在 Hutton，将有机会参加高级统计分析和 R 培训。