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<img src="https://static.igem.org/mediawiki/2019/d/d4/T--Marburg--m_team.jpg" alt="Team"> | <img src="https://static.igem.org/mediawiki/2019/d/d4/T--Marburg--m_team.jpg" alt="Team"> | ||
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<img src="https://static.igem.org/mediawiki/2019/b/b9/T--Marburg--m_logo-text.jpg" alt="Syntex Logo"> | <img src="https://static.igem.org/mediawiki/2019/b/b9/T--Marburg--m_logo-text.jpg" alt="Syntex Logo"> | ||
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With rising atmospheric CO<sub>2</sub> concentrations and declining oil reserves, | With rising atmospheric CO<sub>2</sub> concentrations and declining oil reserves, | ||
we saw that the worldwide effort to change from a petroleum based industry to a carbon neutral industry needs to increase drastically. One | we saw that the worldwide effort to change from a petroleum based industry to a carbon neutral industry needs to increase drastically. One | ||
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<sup>1</sup> Yu, J.; Liberton M.; Cliften, P. F.; Head, R. D.; Jacobs, J. M.; Smith, R. D.; Koppenaal, D. W.; Brand J. J.; Pakrasi, H. B.: | <sup>1</sup> Yu, J.; Liberton M.; Cliften, P. F.; Head, R. D.; Jacobs, J. M.; Smith, R. D.; Koppenaal, D. W.; Brand J. J.; Pakrasi, H. B.: | ||
<i>Synechococcus elongatus</i> UTEX 2973, a | <i>Synechococcus elongatus</i> UTEX 2973, a | ||
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<li><a href="https://www.facebook.com/IGEMMarburg2019/"><img src='https://static.igem.org/mediawiki/2019/b/b1/T--Marburg--m_icon_fa.svg' /></a> | <li><a href="https://www.facebook.com/IGEMMarburg2019/"><img src='https://static.igem.org/mediawiki/2019/b/b1/T--Marburg--m_icon_fa.svg' /></a> | ||
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Revision as of 15:02, 25 July 2019
![Team](https://static.igem.org/mediawiki/2019/d/d4/T--Marburg--m_team.jpg)
![Syntex Logo](https://static.igem.org/mediawiki/2019/b/b9/T--Marburg--m_logo-text.jpg)
Our Project
With rising atmospheric CO2 concentrations and declining oil reserves,
we saw that the worldwide effort to change from a petroleum based industry to a carbon neutral industry needs to increase drastically. One
of the most promising key technologies right now is the use of phototrophic organisms for biotechnological applications. Hence, we decided
quite early this year to devote ourselves to a photosynthetic project. During the design phase, which we initially thought about a project
around the model moss Physcomitrella patens, we soon stumbled upon many common obstacles characteristic to phototrophic chassis due
to our choice of organism. Issues like time intensive culturing and complicated techniques to perform basic molecular biological methods
eventually showed us, why only very few iGEM teams every year decide to use a phototrophic chassis. We saw a need to tackle these issues
and were determined to find a solution.
Consequently, our choice fell on the cyanobacterial strain Synechococcus elongatus UTEX 2973, due to its highly potential doubling
time of about 2 hours 1. This could have a huge impact, as the time consumed by many workflows is mainly dictated by the growth
of your chassis. Our strain could compete with common heterotrophic chassis like yeast, which would be a novelty in photosynthetic
research. We are dedicated to develop this strain as a chassis for the scientific community and future iGEM teams. To restore its natural
competence, which it has lost after isolation, we will integrate a CRISPR/Cpf1 system into our toolbox, enabling easy genomic manipulation
and thus giving us the tools to construct various strains and revert the point mutation responsible for the loss of natural competence.
Additionally, we remove the wild type plasmid pANS from Synechococcus to use it’s origin of replication in our “Marburg Collection
2.0”: a versatile Golden Gate based modular cloning library for fast state of the art assembly of genetic constructs based on a “one step
- one pot” reaction. By designing “operon connectors”, our toolbox is the first to assemble complete operons in the span of two days. This
assembly technique can be performed in our open source liquid handler OT-2 from Opentrons, paving the way for our vision of fully
automated cloning in molecular and synthetic biology - from ordered primers to the finished construct. To add to this vision, we are the
first to establish several laboratory practices in this robot such as colony picking, plating and plasmid purification. By making full
cloning processes possible in the Opentron environment, we give iGEM teams access to an affordable way to accelerate their undertaking,
allowing them to allocate more time to the design of their project. In our metabolic engineering project we prove the value of the tools
we hereby provide: Using our cloning system, we modify our established chassis to produce limonene and farnesene, two valuable
biochemicals that can be used as a biofuel. The chassis’ capabilities are not limited to terpene production but can be expanded to other
areas of biotechnological applications as well as to academic experimental setups. Customized strains offer the opportunity of sustainable
growth in drug development and manufacturing, helping us all to achieve our vision of a more sustainable future on this planet.
1 Yu, J.; Liberton M.; Cliften, P. F.; Head, R. D.; Jacobs, J. M.; Smith, R. D.; Koppenaal, D. W.; Brand J. J.; Pakrasi, H. B.: Synechococcus elongatus UTEX 2973, a fast growing cyanobacterial chassis for biosynthesis using light and CO2. Scientific Reports. 5:8132. DOI: 10.1038/srep08132 (2015)