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We created an easy to use toolbox for <i>Synechococcus elongatus</i> UTEX 2973 to empower rapid design | We created an easy to use toolbox for <i>Synechococcus elongatus</i> UTEX 2973 to empower rapid design |
Revision as of 10:57, 8 December 2019
F A S T E S T .
P H O T O T R O P H I C .
O R G A N I S M .
By providing the fastest growing phototrophic chassis to the community, we are paving the way for other phototrophic organisms in Synthetic Biology.
We created an easy to use toolbox for Synechococcus elongatus UTEX 2973 to empower rapid design testing, including genome engineering tools, self-replicating plasmid systems, natural competence and a Golden Gate-based part library. By providing the fastest growing phototrophic chassis with a doubling time of under 80 minutes to the community, we are paving the way for other phototrophic organisms in Synthetic Biology.
![Strain Engineering Photo](https://static.igem.org/mediawiki/2019/9/91/T--Marburg--hero_strain.jpg)
S T R A I N
E N G I N E E R I N G
We created an "easy to use" phototrophic chassis by restoring the natural competence of S. elongatus UTEX 2973 in order to enormously simplify the transformation process. We established a genome modification system via the CRISPR/Cpf1 and enabled the usage of self-replicating plasmids overcoming the drawbacks of time intensive genome integration for genetic design testing.
![Toolbox Photo](https://static.igem.org/mediawiki/2019/c/ca/T--Marburg--hero_toolbox.jpg)
T O O L B O X
We constructed the green expansion, a set of biobricks to accompany our new chassis. It contains the first MoClo compatible shuttle vector for cyanobacteria. Additionally users can design plasmids for genomic integrations using novel rationally designed integration sites. To improve standardization in phototrophic research we deliver standardized measurement entry vectors to test BioBricks in cyanobacteria.
![Measurement Photo](https://static.igem.org/mediawiki/2019/c/c3/T--Marburg--hero_measurement.jpg)
M E A S U R E M E N T
Following the call for long needed standardization in the cyanobacterial community we ventured out to rationalize important measurements, such as those of light intensities, hugely affecting the growth of our cultures. As using fluorescence for part characterization proves difficult in self-fluorescent cyanobacteria, we showed that the use of bioluminescence reporters offers promising alternatives to improve these characterizations. In order to show that for these measurements clear parameters have to be set, we measured gene expression levels under different conditions using FACS. We additionally employed FACS measurements with accurate cell counts to redefine the way growth curves are done.
![Automation Photo](https://static.igem.org/mediawiki/2019/f/f9/T--Marburg--hero_automation.jpg)
A U T O M A T I O N
The goal of the automation lab was to completely automate the process of cloning using OT-2 Pipetting robots. This was achieved using a state of the art faster-RCNN neural network and a self made camera module for colony picking as well as a self made light table. We also automated large scale purification of plasmids. Our software as well as hardware blueprints are published for the scientific community to give everybody access to scalable and affordable automation.