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                    <p class="lead"> As mentioned previously, our goal was to actively engage the scientific community and the general public to assure that our project is safe, responsible and good for the world. This was among other things made possible through public presentations and active dialogues at events & conferences. A big applause to everyone that made these events possible!<br><br>
  
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You can find a more detailed overview over all the events or conferences that we have organised and participated in below:
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                    <h2 class="section-heading">Introduction </h2>
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                    <p class="lead">Our team believes that establishing a stable platform for scientists to create naïve orthogonal living compartments, would allow for an unpredictable advancement in the field of synthetic biology. Our project will not attempt to create an endosymbiont, but instead investigate the mechanisms in free-living cells in a bottom-up approach to endosymbiosis. 
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The endosymbiotic theory, formulated in the early years of the previous century, outlines that the organelles of the eukaryotic cell, such as the mitochondria, have their origin in free-living prokaryotes engulfed by bigger cells. These incorporated cells then co-evolved with their host conferring to it novel emergent properties which ultimately helped fuel the development of more complex multicellular biological systems such as plants and animals (Archibald, 2015). </p>
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<p>We have identified three mechanisms we believe to be mandatory for the development of a stable endosymbiotic relationship, which we will be trying to replicate in free-living cells. First of all, in order for the relationship to be stable, the two organisms must  be mutually dependent on each other; there must be a mutually beneficial interaction between host and symbiont. Secondly, there has to be some sort of control and synchronization of symbiont replication. If the symbiont were to be replicating freely we could end up with way too many or not enough symbionts in the host.  Finally, a common feature of the endosymbiotic organelles we have looked at, is the transfer of genes from the symbiont to the host. Because of this transfer, the gene and protein expression is taking place in the nucleus and the proteins and metabolites are transported to the organelle. This import of proteins is interesting not just for understanding endosymbiosis, but also for the potential applications in synthetic biology.</p>
 
  
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<p>Based on these considerations, we decided to work on three distinct, but intertwined, projects pertaining to endosymbiosis, namely Interdependence, Number Control, and Protein import. We believe that by combining these three projects, a key step towards the understanding of endosymbiosis and its employment in synthetic biology will be obtained. </p>
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The Nordic iGEM Conference (NiC) is an annual meetup for all nordic teams from Norway, Sweden, Finland and Denmark. <br><br>The 9 -11th of June Incell hosted the conference at the Department of Plant and Environmental Sciences,  in which enriching workshops were held along with a mini-Jamboree to gather feedback on each other’s projects.
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<br><br>On the last night, a gala dinner was organised as a final opportunity for the different Nordic iGEM teams to bond.
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<figcaption><b>Image 1 </b>Will Wright at Nordic iGEM Conference (June, 2017).</figcaption>
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                    <h2 class="section-heading">Applications and Implications</h2>
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<h2 class="section-heading">Lecture Session: Synthetic Biology</h2>
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                    <<p>By understanding the basic principles behind the creation of stable endosymbiotic events we hope that in the future it will be possible to use artificial endosymbiosis as a new technology in synthetic biology, and we believe that value can be created in the foundational track of the iGEM competition. History has shown that great scientific advances has followed the implementation of new revolutionary technologies (Gershon 2003). </p>
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<p class="lead">Synapse - Life Science Connect is a non-profit student organization based in Copenhagen. with focus on bridging the gap between student and the life science industry. <br><br>
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<p>We envision that artificial endosymbiosis could be applied in a broad range of fields, including agriculture, medicine and production of valuable compounds. A deeper understanding of the relationships intertwining endosymbionts and their hosts could unravel new knowledge applicable for the treatment of mitochondrial diseases, while a living compartment able to fixate nitrogen from the air could decrease the fertilizer use in agricultural production. </p>
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On the 27th of September, in collaboration with Synapse, inCell hosted a lecture session on synthetic biology at The Maersk Tower in Copenhagen with the attendance of Associate Professors Rasmus John Normand Frandsen and Christopher Workman from the Technical University of Denmark (DTU), co-founder and CSO of Biosyntia Hans Gene and DTU BioBuilders 2017. <br><br>
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The event introduced the participants to the area of synthetic biology from both an academic and industrial perspective. Moreover, a presentation of the framework of the iGEM competition, its past and future was given, as an introduction to the project presentations of DTU BioBuilders 2017 and Incell.
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<p>However, the applications are only limited by the imagination of future users. Indeed, the game-changing role of endosymbiosis has not gone unseen to the eyes of the modern bioengineers, who predict that the establishment of a novel interaction has the potential to radically alter the host cell physiology without directly affecting the host genome (Scientific America Vol 105 pp. 36-45).</p>
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<figcaption><b>Image 2 </b>Synapse evening with Incell and other students.. </figcaption>
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<p>Before the potential application of artificial endosymbiosis, there are many things to consider. While the current regulations regarding GMO limits what is possible to apply in agriculture and medicine, regulations regarding synthetically modified organisms (SMOs) have not yet been systematically put into place. How will a new field of SMO be regulated, and how will it influence possible applications of artificial endosymbiosis?</p>
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<p>In addition to our scientific investigation we are enthused to trigger debate about synthetic biology. We intend to podcast intriguing conversations with experts, thereby hoping to reach the general public and impel the discussion about the ethics and future prospects in combining biology and engineering.</p>
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                <h2>Scientific approach description</h2>
 
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<p>For both the Interdependence and Number Control projects we have chosen to use Escherichia coli as our chassis. As for the Endocytosis, multiple bacteria spanning different taxonomical groups will be examined. In the following paragraphs, a concise experimental plan for each of the projects will be described.</p>
 
  
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<p>The goal of the interdependence project is to explore the use of amino acid auxotrophy as the framework for a mutually beneficial relationship between two different organisms, namely engineering bacteria to export essential nutrients in a readily available form and in the proper quantity to be sufficient for the host. This will create a strong dependency in a host lacking the production facility of these nutrients. We will perform the experiments on free-living cells, and investigate if the exported amino acids would be sufficient to sustain a S. cerevisiae host. </p>
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<h2 class="section-heading">Vartovs Videnskab - Ethics and Future of Synthetic Biology</h2>
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<p class="lead">Vartovs Videnskab (VV) is a small organization driven by Jamila Lilja and Anna Øhlenschlæger, that arranges evening events on scientific topics for a diverse group of young, old, science nerds, curious minds and entrepreneurs. <br><br>
  
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<p> As part of the interdependence project the following will be examined:</p>
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The 21th of September inCell and Vartovs Videnskab hosted an evening in the sign of synthetic biology for more than 130 individuals.. First, a presentation to Incell’s project was given by team members Jon and Sissel. Subsequently, an insight into synthetic biology’s “green” future was given by Postdoc in plant biochemistry at the University of Copenhagen Thomas Laursen. Thereafter, associate Professor in Philosophy at the University of Copenhagen Sune Holm shared his thoughts on the ethical considerations related to synthetic biology and addressed the need of public discussion of the future of synthetic biology.<br><br>
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<li> the functional expression of amino acid exporters in E. coli; </li>
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<li>the level of amino acid produced and exported by the symbiont;</li>
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<li> the effect of nutrient export on E. coli cell viability and growth; </li>
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<li> Grow auxotrophic S. cerevisiae in co-culture with respective amino acid exporting E. coli. </li>
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<li>In silico modelling growth of S. cerevisiae under the levels of amino acids that can be exported by E. coli to investigate the minimum number of E. coli symbionts to sustain an auxotrophic yeast host cell.</li>
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A panel discussion gave the participants the opportunity to directly raise questions on the future of synthetic biology in regards to, among other things,artificial intelligence, robotics, IP rights and the public GMO debate in Denmark.
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<h3>Number control</h3>
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<p>The goal of the number control project is to lower or stop symbiont replication in case of a high symbiont abundance and/or a starvation status of the host cell, with proof of concept being performed in E. coli cultures. To intertwine the cell replication cycle and lower the stress created by the presence of a symbiont inside the cytoplasm of the host, we aim to put the number control system under control with three signals: the symbiont abundance (a quorum sensing circuit), the host cell starvation status, and the host cell replication. QS and cell starvation should lower symbiont replication, while host replication should increase it.
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We aim to establish a replication control and intertwinement using a modular system based on CRISPR/Cas9 technology. A catalytically-dead Cas9 (dCas9) lacking endonuclease activity and a small guide RNA will be guided via RNA-DNA interaction to the origin of replication on the bacterial chromosome, to efficiently and transiently inhibit the chromosome replication (Wiktor J. et al, 2016).</p>
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<p>The dCas9 system will be put under control of the quorum sensing pathway with the lux promoter, which will be active during high symbiont density.
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Additionally, we aim to put the number control system under control of cell starvation status and host cell replication. Moreover, to overcome an unrestrained cell growth, we aim to inhibit the membrane production silencing the expression of a key enzyme for lipid biosynthesis, i.e. Enoyl Acyl Reductase, commonly target of bacteriostatic drugs.</p>
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<p> As part of the Number Control project the following will be examined:</p>
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<li> The effect of dCas9 expression on the cell cycle of E. coli </li>
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<li>The effect of sgRNA binding to various site of the chromosomal origin of replication  on the cell cycle of E. coli</li>
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<p>The integration of the host signals, i.e. the host starvation status and the cell cycle phase, to control the dCas9 expression will be tested using an in-silico model.</p>
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<p>The goal of the protein import project is to import fluorescent proteins into a bacteria by covalently connecting the protein to a cell-penetrating peptide (CPP).</p>
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<p>CPPs are small peptides (down to 8 amino acids), generally rich in arginine molecules, capable of initiating cellular uptake of a large variety of molecules and proteins by inducing endocytosis. In both plant and mammalian cells CPPs have been shown to mediate protein uptake by both covalent and non-covalent association, with increased specificity during covalent association. However, in bacteria only non-covalent uptake of proteins have so far been demonstrated (Chang et al. 2014). We will explore the utilization of the synthetic CPP nona-arginine (R9) as a vector for facilitation of targeted protein import in bacteria.</p>
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<p> As part of the Endocytosis project the following will be examined: </p>
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<li> An initial screening process aimed at evaluating the ability of a variety of bacterial species to take up fluorescent proteins through covalent and non-covalent CPP association </li>
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<p>Proteins needed for this sub-project will be expressed with His-tags in E. coli and purified using affinity chromatography. Both flow cytometry and confocal microscopy will be used to evaluate bacterial protein uptake through use of the fluorescent proteins YFP and BFP as previously described (Chang et al. 2014). A lipophillic dye capable of staining membrane-derived vesicles (e.g. FM4-64) will be used to investigate cellular localization. To facilitate cleavage of R9 following import, either an endogenous or heterologously expressed peptidase will be used. Successful cleavage will be assessed through use of proteomics (e.g. targeted proteomics). The exact details for evaluation of enzymatic ability following protein import have still not been elucidated, we are however currently contemplating using an enzyme whose activity can be monitored through a colorimetric assay (e.g. 𝜷-lactamase).</p>
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Spreading the knowledge of the synthetic biology is an very important mission for the inCell team. We think that it is particularly important to communicate and discuss synthetic biology with the younger generations and future scientists/practitioners of science. <br><br>
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Therefore, we teamed up with the former iGEM team of University of Copenhagen, Cosmocrops, to communicate the topic of synthetic biology and our respectice iGEM projects to Ungdommens Naturvidenskabelige Forening (UNF) - the association of science for youth, ranging from high school to university.
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<li> Archibald, J. M. Endosymbiosis and Eukaryotic Cell Evolution. Current Biology 2015, 25: 911–921. </li>
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<li>Diane Gershon Technology: pushing the boundaries of scientific discovery. Nature Medicine 2003; 9:97</li>
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<p class="lead">The 30th of August, the 2017  iGEM Team representing the Southern University of Denmark hosted a workshop focusing on the ethical aspects of synthetic biology and good practices for the coding of the wiki. <br><br> This event inspired many insights that we subsequently integrated into our project.
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<li>Microsugar Chang, Yue-Wern Huang, Robert S. Aronstam and Han-Jung Lee; Cellular Delivery of Noncovalently-Associated Macromolecules by Cell-Penetrating Peptides. Current Pharmaceutical Biotechnology 2014; 15: 267-275. </li>
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                     <h2>Find Incell here:</h2>
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Latest revision as of 00:00, 2 November 2017

E V E N T S


As mentioned previously, our goal was to actively engage the scientific community and the general public to assure that our project is safe, responsible and good for the world. This was among other things made possible through public presentations and active dialogues at events & conferences. A big applause to everyone that made these events possible!

You can find a more detailed overview over all the events or conferences that we have organised and participated in below:

Nordic iGEM Conference

The Nordic iGEM Conference (NiC) is an annual meetup for all nordic teams from Norway, Sweden, Finland and Denmark.

The 9 -11th of June Incell hosted the conference at the Department of Plant and Environmental Sciences, in which enriching workshops were held along with a mini-Jamboree to gather feedback on each other’s projects.

On the last night, a gala dinner was organised as a final opportunity for the different Nordic iGEM teams to bond.

Image 1 Will Wright at Nordic iGEM Conference (June, 2017).

Lecture Session: Synthetic Biology

Synapse - Life Science Connect is a non-profit student organization based in Copenhagen. with focus on bridging the gap between student and the life science industry.

On the 27th of September, in collaboration with Synapse, inCell hosted a lecture session on synthetic biology at The Maersk Tower in Copenhagen with the attendance of Associate Professors Rasmus John Normand Frandsen and Christopher Workman from the Technical University of Denmark (DTU), co-founder and CSO of Biosyntia Hans Gene and DTU BioBuilders 2017.

The event introduced the participants to the area of synthetic biology from both an academic and industrial perspective. Moreover, a presentation of the framework of the iGEM competition, its past and future was given, as an introduction to the project presentations of DTU BioBuilders 2017 and Incell.






Image 2 Synapse evening with Incell and other students..

Vartovs Videnskab - Ethics and Future of Synthetic Biology

Vartovs Videnskab (VV) is a small organization driven by Jamila Lilja and Anna Øhlenschlæger, that arranges evening events on scientific topics for a diverse group of young, old, science nerds, curious minds and entrepreneurs.

The 21th of September inCell and Vartovs Videnskab hosted an evening in the sign of synthetic biology for more than 130 individuals.. First, a presentation to Incell’s project was given by team members Jon and Sissel. Subsequently, an insight into synthetic biology’s “green” future was given by Postdoc in plant biochemistry at the University of Copenhagen Thomas Laursen. Thereafter, associate Professor in Philosophy at the University of Copenhagen Sune Holm shared his thoughts on the ethical considerations related to synthetic biology and addressed the need of public discussion of the future of synthetic biology.

A panel discussion gave the participants the opportunity to directly raise questions on the future of synthetic biology in regards to, among other things,artificial intelligence, robotics, IP rights and the public GMO debate in Denmark.





Image 3 Incell at Vartovs Videnskab with Postdoc Thomas Laursen and Sune Holm.

Ungdommens Naturvidenskabelige Forening (UNF)

Spreading the knowledge of the synthetic biology is an very important mission for the inCell team. We think that it is particularly important to communicate and discuss synthetic biology with the younger generations and future scientists/practitioners of science.

Therefore, we teamed up with the former iGEM team of University of Copenhagen, Cosmocrops, to communicate the topic of synthetic biology and our respectice iGEM projects to Ungdommens Naturvidenskabelige Forening (UNF) - the association of science for youth, ranging from high school to university.

Image 4 Incell and Cosmocrops at UNF.

SDU Wiki writing workshop

The 30th of August, the 2017 iGEM Team representing the Southern University of Denmark hosted a workshop focusing on the ethical aspects of synthetic biology and good practices for the coding of the wiki.

This event inspired many insights that we subsequently integrated into our project.

Image 5 DTU BioBuilders, SDU iGEM and Incell

Find Incell here: