Københavns Universitet

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Crispr/Cas9 technology to develop plant-based protein for human consumption at Københavns Universitet

Posted 3 weeks ago

Project description
Towards sustainable development of food for future world population (UN SDG 2), and according to 2019 report from ICPP, climate challenges dictate a change towards plant-based rather than meat-based protein sources in human consumption. Today, the protein-rich seed meal in oil-producing Brassica crops is a by-product with limited use as protein source due to the presence of anti-nutritional factors. The emerging trends for plant-based proteins have potential to increase demand for seed meal from oil-producing Brassica crop. The quality of the amino acid profile is excellent – thus making the seed meal a potential very attractive alternative plant-based protein source. We will use the new CRISPR/Cas9 genome editing technology that has revolutionized breeding efforts with its ability to precisely and effectively target specific sites in genomes.
Perspective
The goal is to increase the value of seed meal of oilseed rape as plant-based protein for human consumption by removal of the toxic and bitter compounds by Crispr/Cas9-based genome editing. This will provide a sustainable, local-grown protein source to replace the import of soybean from North and South America.

Your task (can be adjusted based on interest):

  • design and test of gRNA targeting specific gene in Arabidopsis model plant (proof-of-concept)
  • identify orthologs of specific genes in Brassica napus
  • design and test efficiency of gRNAs targeting the orthologs in rapeseed protoplasts
  • Agrobacterium-mediated transform rapeseed and analysis of impact

Methods used
The project will focus on implementation of CRISPR/Cas9 into a crop. You will design and screen for efficiency of guideRNAs targeting key genes responsible for the accumulation of anti-nutritional factors. The guideRNA will be integrated into constructs to be tested by transformation of constructs into protoplast. The efficiency will be validated using different methods (PCR, sequencing, metabolic profiling). Subsequently, regeneration of protoplast will be performed. In our work, we use molecular cloning, expression analysis, bioinformatics, metabolite profiling, plant tissue culture methodology. The project provides a solid foundation for future work in both research labs and biotech industry, and gives a solid foundation in basic skills in plant biology and biotechnology.

Who we are
Research group of 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).

Main supervisor
Head of Center, Prof. Barbara Ann Halkier, DynaMo Center of Excellence, PLEN, Frederiksberg. Email: bah@plen.ku.dk

Co-supervisor: Assistant Professor Deyang Xu, email: dyxu@plen.ku.dk

Keywords
Plant biotechnology, sustainability, Crispr/Cas9, genome editing, molecular biology, analytical chemistry, plant-based protein, expression analysis, genotyping

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