Team:Marburg
Establishing cell-free systems from chloroplasts as rapid prototyping platforms for plant SynBio
THE CHALLENGE
Climate change presents agriculture with the biggest challenge in the history of
humanity. Higher temperatures, droughts and flooding will cause even greater complications and will further
stress our food supply chain.
THE PROBLEM
To tackle these challenges we need crops, which can withstand all these issues.
But one problem we are currently facing is the speed of innovation in crop breeding and improvement, which is
far too slow. In just 30 cycles of sowing and harvesting, we will reach the critical year of 2050, where
humanity has to feed 10 billion people.
OUR SOLUTION
We have successfully developed cell-free systems of chloroplasts for various plant
model organisms and industrially relevant crops to accelerate the design-build-test-learn cycle of
plant Synthetic Biology. With this technology, we offer a drastic time reduction for the genetic engineering of
different plants.
T. AESTIVUM
Bread wheat is the leading source of vegetable protein and the most planted food crop
in the world with a production of over 765 million metric tons. However, engineering wheat is
difficult and slow. Therefore, we have successfully developed a cell-free system of wheat chloroplast extracts,
which allows high-throughput testing of novel wheat parts.
Q. ROBUR
Climate change is affecting our forests drastically and is causing forest dieback.
Designing, building, or even testing BioBricks for a tree seems impossible in the time frame of an iGEM
project. But imagine it would be possible! After several rounds of the DBTL-cycle, we were able to characterize
the first BioBricks of a tree species.
N. TABACUM
Tobacco is the model chassis for chloroplast engineering as the transformation
protocol is the most developed and efficient known today. Therefore, we decided to include tobacco as a model
for
our cell-free systems in order to characterize our novel parts and compare our in vitro data to actual
in vivo
experiments.
S. OLERACEA
To make our workflow of using cell-free chloroplast systems as accessible as possible
for future iGEM teams, we explored a low resource approach, which would not require any greenhouses or other
plant growing capabilities: We established cell-free systems of spinach, which we bought freshly on the market!
BUT WHY CHLOROPLASTS?
Chloroplasts offer several advantages, such as high precision genetic engineering via
efficient homologous recombination, the absence of transgene silencing, the possibility of transgene stacking in
synthetic operons and the potential for high-level expression of gene products.
IMPROVED BIOCONTAINMENT
Chloroplasts are inherited maternally and therefore are not transmitted via pollen.
Their targeted engineering opens up new dimensions of breeding safer, genetically modified crops.
CHLOROPLAST PART COLLECTION
We have designed, built and extensively characterized a novel part collection, which
contains 154 basic parts, tailored for the use in chloroplasts. We could show that chloroplast parts can be used
for a broad species range and are transferable to many plant chassis of your choice, opening up a whole new
world of possible future iGEM projects.
EXPLORATORY DATA ANALYSIS
Chloroplasts originate from a single endosymbiotic uptake of a cyanobacterium that
happened approximately 1.7 billion years ago. Since then chloroplast genomes have been heavily reduced and
evolution has explored a highly dimensional sequence space. We have created a database of more than 6000
chloroplast genomes, with 2.4 million features, and used this database to train a neural network in order to
design, build and test universally usable chloroplast promoters.
HUMAN PRACTICES
We have reached out to numerous important stakeholders of the agricultural sector,
such as Corteva, KWS, the European food safety authority, the central commission for biological safety (ZKBS)
and many more. We also talked to farmers and discussed the implication of our project on their everyday work.
EDUCATION & COMMUNICATION
As it is almost impossible to bring genetic engineering experiments to a classroom
within Germany, we talked to local authorities, to clarify the legal status of using cell-free systems
in a classroom setting. We then successfully tested and evaluated them in schools and finally organized a long
term agreement with the german association for synthetic biology (GASB) to bring this to german classrooms on a larger
scale in the coming years.
PHOTOTROPH COMMUNITY
Projects using phototropic chassis are highly underrepresented in the iGEM
competition. Compared with more common organisms, working with phototrophs entails many challenges. That
is why, we teamed up with the iGEM Bielefeld team for the entire season with the aim to establish a phototroph
community by creating a platform to connect and communicate. Additionally, we prepared a
160-page handbook, which will help future iGEM Teams to start their own phototrophic project.
IGEM TEAM BIOFOUNDRY
Many iGEM Teams do not have access to expensive lab automation equipment and
therefore do not have the same opportunities to implement high throughput experiments in their projects. Because
of this, we teamed up with the Paris Bettencourt team to develop a workflow for decentralized and remote access
to automation. This includes sample logistics, automation experiments and subsequent quality
control.