Difference between revisions of "Team:Bielefeld-CeBiTec/Results"

 
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This is <b>nanoFactory</b>. A combined system with the goal to clean up mining drainage and produce nano particles.
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This is <b>nanoFactory</b> - a combined system to clean up mining drainage and to produce nanoparticles.
  
 
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                       <img class="figure hundred" src="https://static.igem.org/mediawiki/2018/6/6f/T--Bielefeld-CeBiTec--JZ--Overviewwithtext5ds.png">
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                       <img class="figure hundred" src="https://static.igem.org/mediawiki/2018/8/8f/T--Bielefeld-CeBiTec--cg--Overview.png">
 
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Considering Dual Use issues is the principle of considering in which ways the project can be used and misused. After realizing that we never came in touch with this term in our whole academic education we decided to find out more.
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We analyzed the awareness of Dual Use and Dual Use Research of Concern issues through a nationnal and an international survey.
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Both surveys revealed the lack of unified definitions, insufficient education, and failed science communication as major problems. To improve the situation and to prevent restrictions on free research, we increased awareness through improved
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science communication, appealed to the science community, and provided open source material for education of scientists.
 
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„Any modeling project should be tempered by the morality of laziness.“ (Barnes et. al., 2010).
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Modeling contributed to several project parts. One modeling approach identified lethal metal ion
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We model time until cell death, siRNA/RNAi interaction, ferritin characterization and process control parameters for our cross-flow reactor.  
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concentrations and led to the construction of a ROS reducing system to improve the tolerance towards metal ions.  
 
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Due to the lack of data on precise promoter strength data a new measuring system had to be established.
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Since there is a shortage of  reliable information about promoter strengths, we tested a promoter and RBS library to identifiy the appropriate combination for our project.
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Therefore, we constructed a plasmid backbone, which enables reliable promoter strength measurement through normalization based on a second reporter encoded in the backbone. It has not escaped our notice that this system could be applied by further iGEM teams to characterize any promoter sequence of interest.
 
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<h2><a href="https://2018.igem.org/Team:Bielefeld-CeBiTec/Toxicity_Theory">Toxicity</a></h2>
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<h2><a href="https://2018.igem.org/Team:Bielefeld-CeBiTec/Toxicity_Results">Toxicity</a></h2>
  
 
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Metal ions cause hydrogen peroxide to form toxic reactive oxygen species. To counteract, new H<sub>2</sub>O<sub>2</sub> scavenging BioBricks were needed.
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Metal ions have a toxic effect on <i>Escherichia coli</i> cells. We identified critical concentrations for our experiments and developed several methods to reduce ROS.
 
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<h2><a href="https://2018.igem.org/Team:Bielefeld-CeBiTec/Accumulation">Accumulation</a></h2>
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<h2><a href="https://2018.igem.org/Team:Bielefeld-CeBiTec/Accumulation_Results">Accumulation</a></h2>
  
 
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Metal ions need transport proteins to enter the cells. New BioBricks are required to increase the uptake of metal ions to increase the nanoparticle yield.
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To increase the nanoparticle yield, we cloned and characterized dedicated metal ions importers. We investigated the specifcity towards their respective ions and the influence on the growth.
 
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<h2><a href="https://2018.igem.org/Team:Bielefeld-CeBiTec/siRNA">Silencing</a></h2>
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<h2><a href="https://2018.igem.org/Team:Bielefeld-CeBiTec/siRNA_Results">Silencing</a></h2>
  
 
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Knocking out copper exporters causes cell death. Precise control over the translation process leads to a better growth condition for the cell.
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We designed and assembled vectors for assessment and expression of siRNAs. We used our software to design suitable siRNAs and developed an improved vector set for experimenntal validation.
 
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<h2><a href="https://2018.igem.org/Team:Bielefeld-CeBiTec/Ferritin">Nanoparticles</a></h2>
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<h2><a href="https://2018.igem.org/Team:Bielefeld-CeBiTec/Ferritin_Results">Nanoparticles</a></h2>
  
 
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Nanoparticles are materials on nano scale. They show different physical properties and thus, are of high interest for industry, medicine and science.
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We were able to enhance iron nanoparticle formation by overexpressing ferritin in <i>Escherichia coli</i>. Furthermore, we developed a mutated variant of the human ferritin to produce gold and silver nanoparticles.
 
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To filter high amounts of mining drainage without releasing GMO into the environment a hardware need to be designed, constructed and optimized.
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We designed and printed a customized cross-flow bioreactor to filter huge amounts of mining drainage while accumulating metal ions. Through iterated feedback from our modeling we improved our prototype and developed an improved bioreactor to facilitate application of our system for the cleaning of mining drainage.
 
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Different applications for nanoparticles received through synthetic biology are investigated.
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During our project we were able to demonstrate accumulation of metal ions in <i>Escherichia coli</i>, while increasing the tolerance towards such ions. We engineered ferritin to enable iron, silver and gold nanoparticle formation. Furthermore, we demonstarted that nanoparticles could be used to print conductive paths.
 
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Latest revision as of 04:23, 6 December 2018

Results Overview
This is nanoFactory - a combined system to clean up mining drainage and to produce nanoparticles.

Dual Use

We analyzed the awareness of Dual Use and Dual Use Research of Concern issues through a nationnal and an international survey. Both surveys revealed the lack of unified definitions, insufficient education, and failed science communication as major problems. To improve the situation and to prevent restrictions on free research, we increased awareness through improved science communication, appealed to the science community, and provided open source material for education of scientists.

Modeling

Modeling contributed to several project parts. One modeling approach identified lethal metal ion concentrations and led to the construction of a ROS reducing system to improve the tolerance towards metal ions.

Promoter Collection

Since there is a shortage of reliable information about promoter strengths, we tested a promoter and RBS library to identifiy the appropriate combination for our project. Therefore, we constructed a plasmid backbone, which enables reliable promoter strength measurement through normalization based on a second reporter encoded in the backbone. It has not escaped our notice that this system could be applied by further iGEM teams to characterize any promoter sequence of interest.

Toxicity

Metal ions have a toxic effect on Escherichia coli cells. We identified critical concentrations for our experiments and developed several methods to reduce ROS.

Accumulation

To increase the nanoparticle yield, we cloned and characterized dedicated metal ions importers. We investigated the specifcity towards their respective ions and the influence on the growth.

Silencing

We designed and assembled vectors for assessment and expression of siRNAs. We used our software to design suitable siRNAs and developed an improved vector set for experimenntal validation.

Nanoparticles

We were able to enhance iron nanoparticle formation by overexpressing ferritin in Escherichia coli. Furthermore, we developed a mutated variant of the human ferritin to produce gold and silver nanoparticles.

Reactor

We designed and printed a customized cross-flow bioreactor to filter huge amounts of mining drainage while accumulating metal ions. Through iterated feedback from our modeling we improved our prototype and developed an improved bioreactor to facilitate application of our system for the cleaning of mining drainage.

Proof of Concept

During our project we were able to demonstrate accumulation of metal ions in Escherichia coli, while increasing the tolerance towards such ions. We engineered ferritin to enable iron, silver and gold nanoparticle formation. Furthermore, we demonstarted that nanoparticles could be used to print conductive paths.