Expanding the range of barley and potato gene editing tools.

培育新的高产、安全、营养、气候适应性强的作物，对于缓解农业面临的许多有据可查的压力至关重要。基因编辑(GE)或精确育种是一种从根本上不同于传统育种和转基因方法的显着工具。在农作物物种中，可能潜在证明社会、环境和经济价值的适当基因靶标仍然很少。学生将探索大麦和土豆中的转基因目标，提供适当的背景和理由来解释为什么这些目标可能是重要的。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，以改善大麦和马铃薯的表现。选定的关键基因靶点将用于实际示范GE项目。利用已建立的CRISPR/CAS9系统，我们在产生大麦编辑品系方面取得了一些成功。可以使用我们已有的方法创建新的样本基因靶点。然而，GE是一种新兴和快速发展的技术，使用碱基编辑或通过碱基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，将A/T转换为G/C。初级编辑系统包括使用Cas9尼克酶-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1、2、3组成的pegRNA。学生身份将允许在我们的作物中建立和发展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，创造出含有CRISPR-Cas9成分的转基因植物，以便发生突变。有一些方法可以完全消除这一中间步骤，方法是通过将组装好的Cas9蛋白-gRNA瞬时传递到细胞中来产生突变，通常是通过传递到植物原生质体。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。但在从最合适的组织中分离出原生质体以及从原生质体中再生植物方面，原生质体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输送方法。转基因技术的应用在欧洲仍然存在争议，并受到从公众到科学界和政策制定者的各级辩论。这是参与这场辩论并探索通用电气的社会、环境和营养益处的理想方式。该奖学金将在赫顿建立的生物技术设施中提供GE方法学方面的培训，并配备具有植物转化经验的工作人员。主管与在该领域工作的其他团队有良好的联系，并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。学生将拥有通过伊斯特比奥、邓迪大学和赫顿参加多种培训机会的优势。例如，在赫顿，将有机会参加高级统计分析和R培训。

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.