Team:Heidelberg LSL/Project Summary

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Summary & Outlook

Lying in the sun we now have time to recover and think about the past weeks…

Summary

After a long phase of planning and development, we sucessfully developed a new biological system in E.coli that reacts quantitatively to UV-radiation in both artificial, and natural doses of radiation.

All of our promoters, precA, precB and psulA, which are linked with the SOS-response, reacted reliably to UV-induced damages and started exprimation of our reporter genes GFP and LacZ. All combinations such as precA_LacZ, precA_GFP, precB_LacZ, psulA_LacZ and psulA_GFP worked regarding quantitative UV-detection, which is a great surprise and unpredictable success. Outdoor tests with our placZ-reported constructs in direct sunlight and in shade indicated a light-dependent color change which is visible even to the naked eye. The color of the exposed bacterial suspension changed in the course of expose from light-yellow to a dark greenish blue. This color change is a result of the UV-induced exprimation of the enzyme lactase which caltalyses the reaction of XGal to a colored product. With our test series, we furthermore discovered significant differences in the strength of UV-detection between rec- and sulA-promotors. precA_LacZ constructs for instance showed less visible color change after 8 minutes of sun expose than psulA_LacZ constructs.
These successful real life tests give evidence to the possibility to use our biological systems in a real applicational context such as our iGEM-jewelry collection.

After these successful construtions and tests we began to characterize carefully our new biological systems and submitted four new entries to the parts registry. To make this important contribution to the synthetic biology community and to the partsregistry, we adjusted our constructs to the official requirements of the partsregistry such as clone it into the pSB1C3 plasmid backbone with the required pre- and suffix. Our new three parts are precA_LacZ, precB_LacZ and psulA_LacZ. Additionally, we looked up, ordered, tested and registered the sequence of the recB- promotor, which was not yet available in the registry. First tests showed that UV-induced activity of precB can be compared with those of precA.

With this unpredictable success, we are glad to be able to bed this new and innovative UV-detecting system in a fancy application: iGEMS. iGEMS are small tubes filled with bacterial suspension and trailed to trendy necklaces and bracelets. We developed this fictitious product in order to raise the public awareness of the “invisible danger”, to pique the reader’s curiosity and to present our project in an easy-to-understand way. With the establishment of a fictitious ‘Online Shop’ we tried to put genetically modified organisms in a modern and familiar background. As a open-minded part of the society, this should encourage the reader to think again about synthetic biology and to begin controversial discussions about this upcoming field of science.
For us personally, the project broadened our scientific horizon and gave us the opportunity to learn how to work scientifically. We had a great time together and look forward to present our work to you at the Jamboree 2012.

Outlook

In the past few month the project was brought up to a great level. We have constructed different BioBricks which were sent to the partsregistry, tested our parts and designed a jewelry collection. But furthermore we have found difficulties that should be optimised in the future. We have worked with enthusiasm on the bacteria tool kit which can detect UV-radiation and have made a lot of progress, therefore we want to continue working on it.

In the future we want to create a clearer characterisation of different members of the same gene family as precA. So Our assumption is that different rec genes are activated based on a certain characteristic extension of DNA damage dependent on UV radiation. By linking these genes together we want to generate a more gradual UV-response for the detection of qualitative UV-intensity. At UV-intensities different genes are activated and a color gradient appears. This gradient will be used to determine the exact amount of radiation. This is important to give the bacteria tool kit the properties of a dosimeter. We started testing precB and precC, they already gave us a positive result, so that our assumption was right. Now we have to analyse the data to find their most effective use.

For the iGEM competition we only tested our constructs by exposing the samples to UV light. We have not analysed if the construct would also work with radioactive radiation. Radioactive radiation causes different DNA damage, therefore it is not sure if precA becomes activated in the same way as with UV radiation. In principle, we assume that the induction by radioactive radiation would work as the precA promoter is known to be activated by the induced damage. We could not test our constructs with radioactive radiation, because we did not have the right equipment. But we have got a contact person who could give us access to the required machines.

We also want to include the constructs with a GFP reporter instead of LacZ in our iGEMs collection. GFP cannot be seen by a color change of the bacterial suspension and normally you need further measurement systems to detect GFP, because it has to be excited by a specific wavelength. To overcome this challenge, we want to combine the iGEMS with LED lamps. LEDs emit light of a certain wavelength, e.g. blue light with 460 nm. Therefore we can make GFP visible using blue LEDs. Furthermore we could also develop a construct with dtTomato, a very light, red-orange fluorescent protein, as another reporter. We could generate a color range from green to orange to red with just one single blue LED.

For the production of the jewelry, which shall be once a common product for the public, we need to simplify this system. Only the rec gene is needed that gives us the best adaptation to the human body. The bacterial suspension shall turn blue, when it is irradiated by too much sunlight, that is dangerous for the human. That is a really easy way to recognize UV-radiation, which does not need any further background in synthetic biology. At the moment our bacteria turn blue after 10 min exposure. But the human body can resist much more UV-light without getting DNA-damage. We have to find a rec gene that show the most similarities with the human body. An other possibility to handle this problem would be to vary the UV permeability of the bacteria containment.
Some more problems that came out while working on the constructs. At the moment we have to add X-Gal after the exposure to UV, because UV-radiation may degrade X-Gal. But when producing locked vial, X-Gal cannot added additional. Therefore we have to find by researching different ways to deal with X-Gal. Furthermore we have to take account of the use of suncream in connection with our iGEMs, because suncream delays the effect of UV-light. We are optimistic that we can find solutions to all problems and develop our project to a industrial standard.
Our project comprises a novel idea and opens up new perspectives, especially in Human Practices. The integration of the bacteria into jewelry represents a completely new and consumer-friendly solution to give the public an understanding of synthetic biology. We found an easy way for everyone to protect themselves from sunburn. The method to use dosimeters is old-fashioned, complicated and expensive. Lying on the beach, nobody uses dosimeters because they are too big and uncomfortable. Not only do our fancy iGEMS warn people of too intense UV-radiation but also are trendy accessories! People who wear our iGEMS are aware of the invisible danger and encourage other people to follow this upcoming trend.
In cooperation with the biotechnological companies and the jewelry industry we could develop a new branch of sustainable applications for biological systems.

We are looking forward to the iGem HS Jamboree 2012 and present our work to the other teams from all over the world. We hope to get in contact with other biology interested pupils and present them our iGEMS.

Achievements

Three different UV-Sensor parts constructed and submitted to the registry
All parts were characterized carefully and show a UV-dose-dependent color-production
Outdoor-Test shows our parts to be working in a real application context
Human Practices - consumer product development for raising the public awareness of the “invisible danger” and presenting our project in an easy-to-understand way
Finally, we learned a lot about synthetic biology, laboratory methods and had a great time working together on our project, and are now looking forward to meeting the other teams at the iGEM HS Jamboree!