Expanding the range of barley and potato gene editing tools.

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.