Difference between revisions of "Team:Marburg/Demonstrate"

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Great things are not done by impulse, but by a series of small things brought together. – Vincent van Gogh
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This year we expanded the Marburg Collection from 2018 with 55 new parts to the Marburg Collection 2.0. With our developed workflow we could characterize our parts (Verlinkung) and compare them with a second measurement method: FACs. We added two new features for genome engineering of cyanobacteria: a CRISPR/cpf1 (Link to crispr) guided knockout system as well as a modularized assembly of repair templates for the knock in of genes (M.E.G.A. expansion). This includes integration sites that target conventional neutral sites in cyanobacteria but we also rationally designed two novel integration sites based on RNA-seq data. Additionally we offer the first MoClo compatible shuttle vector for cyanobacteria (Link to best part) and characterized gene expression based on that origin of replication. We used our new shuttle vector to build standardized devices for the characterization of BioBricks in cyanobacterial chassis to improve the reproducibility of results and to simplify large scale assemblies. For this we used placeholders, a novel part type that aids in the construction of a larger set of parts by reducing the involved cost and workload significantly. Additionally we tested our toolbox with PCC 7942 to show that the Marburg Collection 2.0 is also working with similar cyanobacterias. We offer free access to the data of our characterization, enabling the iGEM community and scientists to choose the parts based on this data. To improve the measurement method applicable for cyanobacteria we focused on measurements via luminescence reporters over fluorescence reporters, because of the fact that cyanobacteria emmit autofluorescence. This way our results are way more accurate, because of the reduced background noise. The higher accuracy is obviously visible during the measurement of our parts, where we could see a difference of 5x10^5 between the background noise and the signal, which implements that already a small amount of sample has a more intensive signal.
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Hereby, we want to encourage the community of young scientists to work with the fastest phototrophic organism Synechococcus elongatus UTEX 2973 because of its high relevance for biotechnological applications.
  
  

Revision as of 00:42, 22 October 2019

Wiki Text: Demonstrate “Started from the bottom, now we here” - Drake Our project has three goals. First we want to establish the cyanobacteria Synechococcus Elongatus UTEX 2973 and restore its natural competence. Secondly, we wanted to show the best growing conditions with the goal of standardization for working with UTEX 2973. The final goal is to create a suitable golden-gate toolbox for our strain and share it with other iGEM teams and researchers. In our iGEM year we achieved to restore the natural competence of Synechococcus Elongatus UTEX 2973, analyzed different cultivation parameters (link to growth curves) to accelerate the growth speed including finding the best conditions. Besides that we expanded the Marburg Collection from 2018 and are able to deliver a toolbox of xxx parts with free access to the world. On top of that we automated plasmid purification (link to labautomation) and colony picking via the Opentron OT-2. Great things are not done by impulse, but by a series of small things brought together. – Vincent van Gogh This year we expanded the Marburg Collection from 2018 with 55 new parts to the Marburg Collection 2.0. With our developed workflow we could characterize our parts (Verlinkung) and compare them with a second measurement method: FACs. We added two new features for genome engineering of cyanobacteria: a CRISPR/cpf1 (Link to crispr) guided knockout system as well as a modularized assembly of repair templates for the knock in of genes (M.E.G.A. expansion). This includes integration sites that target conventional neutral sites in cyanobacteria but we also rationally designed two novel integration sites based on RNA-seq data. Additionally we offer the first MoClo compatible shuttle vector for cyanobacteria (Link to best part) and characterized gene expression based on that origin of replication. We used our new shuttle vector to build standardized devices for the characterization of BioBricks in cyanobacterial chassis to improve the reproducibility of results and to simplify large scale assemblies. For this we used placeholders, a novel part type that aids in the construction of a larger set of parts by reducing the involved cost and workload significantly. Additionally we tested our toolbox with PCC 7942 to show that the Marburg Collection 2.0 is also working with similar cyanobacterias. We offer free access to the data of our characterization, enabling the iGEM community and scientists to choose the parts based on this data. To improve the measurement method applicable for cyanobacteria we focused on measurements via luminescence reporters over fluorescence reporters, because of the fact that cyanobacteria emmit autofluorescence. This way our results are way more accurate, because of the reduced background noise. The higher accuracy is obviously visible during the measurement of our parts, where we could see a difference of 5x10^5 between the background noise and the signal, which implements that already a small amount of sample has a more intensive signal. Hereby, we want to encourage the community of young scientists to work with the fastest phototrophic organism Synechococcus elongatus UTEX 2973 because of its high relevance for biotechnological applications.

Demonstrate

Gold Medal Criterion #4

Teams that can show their system working under real world conditions are usually good at impressing the judges in iGEM. To achieve gold medal criterion #4, convince the judges that your engineered system works.

Your engineered system has to work under realistic conditions. Your system must comply with all rules and policies approved by the iGEM Safety Committee. Your system can derive from or make functional a previous iGEM project by your team or by another team. For multi-component projects, the judges may consider the function of individual components.

To be eligible for this award, you must add clear documentation to this page and delete the alert box at the top of this page.

Please see the 2019 Medals Page for more information.