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Transporter engineering to prevent toxins in seeds using Crispr technology

Main area:
Target group:Biology-Biotechnology, Natural Resources, Other Education
Educational level:Master, Project in practice, Project outside course scope
Project description:

Towards sustainable development of food for future world population (UN SDG 2), and according to 2019 report from UN climate panel IPCC, climate challenges dictate a change towards plant-based rather than meat-based protein sources in human consumption. Rapeseed (Brassica napus) is the world’s third largest oil crop and grown for its high-value oil for human consumption and biofuel. Despite much breeding effort, the protein-rich seed meal with an excellent amino acid profile has limited use due to the presence of anti-nutritional factors. Glucosinolates are stored to high concentrations in the seeds, although the seeds have no de novo synthesis of glucosinolates. The goal of this project is to identify and characterize the transporter that is key for driving toxic glucosinolate defense compounds into the embryos in the seed.

Perspective
Understanding this process will advance basic knowledge and plant biotechnology towards plant-based protein for human consumption in a sustainable agriculture.  

Your task (can be adjusted based on interest):
a)     Screen glucosinolate candidate transporter library in yeast and Xenopus laevis oocytes
b)     Biochemical characterization of transporter in yeast (substrate specificity, Km value) and in planta (localization, expression level)
c)     Crispr-based genome-editing and assessment of impact by metabolite profiling and bioassay

Who we are: Research group of Head of Center, Professor Barbara Ann Halkier in the DynaMo Center of Excellence (https://dynamo.ku.dk). We provide a stimulating, international research environment with lots of opportunities to advance your skills. Our research lab has a strong international research profile with a recent record of high impact publications.

Expectations: We expect you to be enthusiastic, dedicated and eager to learn and advance your skills in many disciplines (e.g. research, oral and written presentations).

Methods used:Bioinformatics on candidate gene families, molecular biology, biochemical characterization of transporter(s) in yeast and plant hosts, LC-MS analytical chemistry, design of gRNA, Crispr-based transport engineering, plant genotyping. The project provides a solid foundation for future work in both research labs and biotech industry by advancing skills in Crispr technology, chemical analytics and heterologous expression in several host organisms.
Keywords:Seed loading, transporter characterization, Crispr technology, sustainability
Project home page: https://dynamo.ku.dk/
Supervisor(s): Prof. Barbara Ann Halkier, Assistant Professor Deyang Xu (dyxu@plen.ku.dk)
Email:bah@plen.ku.dk