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.

生产新的高产、安全、营养、气候适应性强的作物，对于减轻农业面临的许多有据可查的压力至关重要。基因编辑（GE）或精确育种是一种显著的工具，与传统育种和转基因方法有着根本的不同。在作物物种中，可能显示出社会、环境和经济价值的适当遗传目标仍然很少。学生将探索大麦和马铃薯中的转基因目标，提供适当的背景和理由来解释为什么这些目标很重要。结果将是一份基因清单，这些基因可以通过精确育种来改变，以提高大麦和马铃薯的性能。选定的关键基因靶点将用于实际的示范GE项目。我们已经在使用已建立的CRISPR/Cas9系统产生大麦编辑系方面取得了一些成功。可以使用我们建立的方法创建新的示例性基因靶标。然而，GE是一种新兴且快速发展的技术，并且使用碱基编辑或通过引物编辑产生任何碱基取代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C/G转换为T/A和A/T转换为G/C取代。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。该奖学金将允许在我们的作物中建立和开发这些新的最先进的编辑程序。在植物中，可以有一个中间步骤来产生含有CRISPR-Cas9组分的转基因植物，以允许突变发生。存在通过使用组装的Cas9蛋白-gRNA到细胞中的瞬时递送来完全去除该中间步骤以产生突变的方法，通常通过递送到植物原生质体。这在直接编辑作物优良品系而不需要转化方面具有很大的优势。但是，原生质体在从最合适的组织中分离原生质体和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辩论。这个奖学金是一个理想的方式来参与这场辩论，并探讨社会，环境和营养的好处转基因。该奖学金将在赫顿已建立的生物技术设施中提供GE方法学方面的培训，并配备有植物转化经验的工作人员。主管人员与从事这一领域工作的其他团队有着良好的联系，可能有机会在其他机构接受相关培训。博士项目将为学生提供分子生物学，显微镜，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个培训机会的优势。例如，在赫顿，将有机会参加高级统计分析和R培训。