CHIC project at the science programme FUTURIS

On Monday, 14th October 2019, the science programme FUTURIS, from the Euronews channel, made a documentary about the CHIC project.

They made several interviews to some of our partners and they also explained the benefits of implementing new plant breeding techniques, in chicory breeding, to convert it into a multipurpose crop and to obtain high value products for consumers’ health benefits.  

On the other hand, our project coordinator, Dirk Bosch had a special interview in the Euronews Bonus Section. He provided detailed information about why we selected chicory as a promising and a strong crop to be converted into a multipurpose one.  

The documentary has been translated into 12 languages and we want to thank the FUTURIS team for guiding us during the filming process and for bringing such a nice result.

Enjoy the documentary!


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CHIC at the European Researchers’ Night in Indjija (Serbia)

IBISS, partner in CHIC Project worked on organizing the presentation regarding the results of our CHIC Project, though team member Jovana Petrovic directly participated in this manifestation.

On September 27th 2019, Jovana Petrovic from University of Belgrade, Institute for Biological Research “Siniša Stanković, National Institute of Republic of Serbia presented knowledge gained so far on health-beneficial effects of Cichorium intybus and its possibilities to be used on every day basis.

People seemed very keen to get to know with facts how this plant that grows on meadows nearby, could exert useful effects as a tonic, coffee substitute and prebiotic.

Since this manifestation is dedicated to popularization of science making it interesting to children of school age, retired people and overall families with children, this Researchers’ Night was a unique opportunity for people to get familiar with new breakthroughs in research that they could directly benefit on.

The presentation of the Project took place at Indjija, the intersection city between Belgrade and Novi Sad.

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Development of four conceptually different New Plant Breeding Techniques

The objective of CHIC’s in this area is to develop four different methods for genome editing of chicory. These methods differ in their degree of ‘DNA invasiveness’: the extent to which CRISPR genes are introduced into the plant’s DNA. In the end all four methods lead to identical genetic outcomes.

CHIC methods  related to the presence of CRISPR DNA in the chicory plant’s DNA

These two methods are based on stable integration of CRISPR genes in the chicory genome, leading to so called transgenic  chicory plants. After the CRISPR genes have done their work, the genes are removed from the genome. In the 1st method the crossing and selection will be used to select offspring plants without the CRISPR genes and with the desired mutation. In method 2, specialized enzymes will be used to remove the CRISPR genes from the plant’s DNA, so making crosses is not needed.

During the 1st two years of the project, thanks to applying the 1st method we have obtained transgenic chicory plants. These plants are now being analyzed for functional edits in the chosen target genes: genes involved in the production of the enzyme germacrene A synthase. This enzyme is essential for the production of terpenes.

Methods related to the non-presence of CRISPR genes in the plant’s DNA

These two methods are based on administering CRISPR tools to chicory cells without incorporating CRISPR genes in the plant’s DNA.

In method 3 plasmid-DNA that harbours CRISPR genes, is introduced into individual chicory cells. Plasmid DNA is best known from bacteria: circular DNA molecules that encode important traits. After introduction into a chicory cell, the CRISPR DNA is expressed and the CRISPR tools are being assembled in the cell. The plasmid DNA is eventually degraded. 

In method 4 the CRISPR tools, protein and guiding RNA, are assembled outside the plant cell and then introduced into plant cells. This way there is no CRISPR DNA entering the plant cells.

For all methods we need to treat individual chicory cells (protoplast). The challenge then is to grow complete plants from these single cells. We have shown that this is indeed possible. We have also optimized techniques to introduce plasmid DNA and the CRISPR tools into chicory cells.

This has already resulted in plant cells in which the genes involved in the production of the enzyme germacrene A synthase are permanently disabled, thanks to the small alteration the CRISPR tools made in the plant DNA.

Optimizing the CRISPR tools

One of the CRISPR tools is a guide RNA (gRNA) molecule. The other is an enzyme called Cas9. The gRNA guides the Cas9 enzyme to the place in the plants’ DNA where a mutation is desired. We have designed gRNAs and have shown that they indeed guide the enzyme, enabling it to temporarily break the DNA at the correct place.

We have obtained plants with desired mutations in both chromosome sets (chicory is diploid) and in many or even all the gene copies chicory possesses. We are now testing the plants for the terpene content that may have some type of bioactivity.

Genome insight

The existing DNA screening techniques are not effective to screen plant material for small changes in the DNA in a high throughput manner. Therefore we are developing a new technique, based on techniques one of the partners developed for kiwi.

During 2019, the partners decided to jointly invest into a better assembled genome sequence of chicory, as the available genomic databases were of insufficient quality for our research.

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