Team:Groningen/Safety

Safety proposal

1.

For the realization of our project we will use the E. coli K-12 derivative DH5α, which is a debilitated strain of the pathogenic wild type E. coli. It lacks cell wall components important for recognition and adherence to intestinal epithelial cells and is hence not able to colonize the intestinal tract of neither humans nor animals. The cultivation of E. coli K-12 derivative strains under laboratory conditions further decreases its ability to colonize the intestine and they are, additionally, easily excluded by the microbiota present in the human colon. Furthermore, E. coli K-12 derivatives are susceptible to stomach acid and can survive it only when introduced in basically buffered media. It does not appear to produce significant amounts of toxins and lacks also other virulence factors. The likelihood of reversion to a pathogenic strain is very low, since E. coli K-12 is a deletion mutant. There is currently no data available which suggests adverse effects of E. coli K-12 on plants or on other microorganisms in the environment. To sum up, E. coli K-12 has a long history of safe use (over 80 years), it has been shown to be avirulent, and the genetic lesions are well understood. It is therefore assigned to the lowest hazard group 1 and can be handled safely at Containment Level 1. None of the sequences we plan to insert is known to be toxic or to possess any other harmful properties to neither human/animal health nor the environment. The engineered organism does not secrete nor produce any chemically active products. The used vectors are without exception non-conjugative, which reduces the chance of horizontal transfer of antibiotic resistances. Thus, our engineered organism will not exhibit any additional hazardous properties to the host organism. The engineered organism is designed for laboratory usage only. No release into the environment or exposure to people is required for the realization of our project. Although we apply good laboratory practice during the work in our lab, we may assume a small chance of unintended release of the engineered organism into the environment. Because of the above mentioned reasons we do not expect any adverse effects on environment or public. In case of changes of function of one or several bioparts due to mutation, we can foresee two scenarios: First, as the working of our system depends on fine tuned parameters, a change would most likely prevent the whole system from working at all. Second, a mutation could cause a continuous activation of one of the output signals. As our output signals are represented by non-toxic proteins we also do not expect any adverse effects in this situation.

2.

All sequences which will be inserted into the host organism are already registered BioBricks. Nevertheless, we intend to make new BioBrick devices which assemble known Biobricks into auto-inducing loop devices. As the involved proteins do not exhibit toxicity or other hazardous properties, our new BioBrick devices do not raise any safety issues. As we plan to make the input, as well as the output signal exchangeable, it is still important to assess which events will lead, for example, to an excessive production of the output signal. Especially, if the non-hazardous proteins we will use, are in case of malicious misuse by third parties, exchanged for more critical proteins, in terms of safety. For this purpose we will try to implement an in silico simulation that will help us to identify the weak points in our system, and to prevent unlikely unexpected scenarios.

3.

At the University of Groningen a biological safety manual, containing safety measures for activities with genetically modified organisms, was composed by the biological safety officers (see http://molgen.biol.rug.nl/molgen/files/VMT.pdf). We had the opportunity to talk to one of these biological safety officer of our University about the potential safety issues of our project. The biosafety-risk level of our proposed project was assessed as minimal. However, we have to take care that all the handlings and storage of the GMOs are carried out at the proper containment level (ML-1) and in the proper labs (ML-1 labs). All team members, who actually perform experiments in the lab, have received a instructions and training how to work in a biosafe manner. Additionally, our lab-work is supervised by a VMT-qualified university employeem who is officially qualified to work with GMOs. The national biosafety regulations and guideline of The Netherlands can be found on the website of the "Commissie Genetische Modificatie" (http://www.cogem.net/).
As we are working with a non-infectious host organism, none of the inserts possesses hazardous properties, and we will use only non-conjugative vectors, applying good laboratory practice and fulfilling Containment Level 1 will be sufficient to minimize the risks for both us, the researches, and the environment.

4.

We collected some opinions within our group on how we think synthetic biology, and especially the iGEM competition, is percieved by the public.

"I think, it depends on the target group you contact. In my personal experience, food companies do not want to have anyting to do with us. The bad publicity of GMOs towards the consumer is a strong driving force. Research focused enterprises like the idea of people working in this field. And it is possible to extrapolate this to the common public. Maybe we should focuse the human practices in informing the general public, as well as the schools."
"I think people do not trust genetic engineering in general. Most of the researches believe that the public has this negative approach because of a deficit of knowledge about the process and its working. On the other hand, people do not have negative approach to computers, cars, planes or microwaves. They do not fear each time they go into a lift that it will kill them, although it could. And most of the people do not know how those potentially deadly devices work - they just trust the manufacturers that they have done a good job. So it is highly possible that the problem is not that people have not enough knowledge, it is that they do not trust the scientific community. This is why all risk assesment procedures, and all procedures in general should be very clear, very systematized and very transparent and this is why iGEM should focus more on developing standards, then on popularizing the knowledge among the society."
"I guess people do not know about it. Most people would not understand when told about iGEM anyway. So I think they position is mostly governed by how they feel about iGEM. If they would learn about it the way they feel about it would be mosly determined by in which context they learned about it. If they hear about it from someone they like, they will like it. If a politician tells them about iGEM they will not like it. In this sence iGEM is prone to generate more controversy and problems as it gets more well known. Staying smaller longer will work in its advantage on the PR side."
"People don't really know what genetic manipulation is and that it can be useful. It has a negative twist in most cases. I think that if the society knew more about it, that it does not need to be harmful for people and that it actually can have great purposes, it can change their mind. You should start at high schools with giving more information in the biology subject, because now I think there is not much told in the biology lessons about synthetic biology. Also tell stuff to people that interests them, like a stay at home mum needs to hear another aspect of synthetic biology than a salesman for computers. The most important thing is to let people trust the biology in synthetic biology. They should know about the safety issues and what protocols are needed to work correctly in the lab."
"In genetic engineering, researchers have been playing around with genetic materials, it certainly raises negative issues to society even it's highly useful, because they thought, researchers have been modifying natural living things, which nowadays society puts a high concern on natural things. Moreover, synthetic biology is a nascent-field, which is designing artificial genetic circuit would be definitely hard to be perceived by public. Since iGEM competition is intended for undergraduate students who want to spend time working in the lab during summer holiday, I think public opinion has a positive response and an enthusiastic support, apart from synthetic biology safety issues."
"I don`t think the general public is interested in understanding complicated things (I don`t exclude myself here!). I actually think it`s important to convince politicians that biological engeneering isn`t a bad thing, but will improve everybodies live, so the politicians can deal with the public. The meeting of minds might be a nice approach..."
"As with all other past technologies where the possibilities were grand, so is the accompanied fear. People and media tend to speculate about worst case scenarios all the time, it's maybe even an useful build in defense mechanism. Attempting to foil that behavioral pattern with insight and understanding is, in my opinion, as noble as it is futile. Therefore my suggestion would be not to focus on the perception of the masses, and keep the iGEM competition active and visible within its own scene and not beyond that. Only time and dependence upon it's products will make synthetic biology an acceptable factor in our risk society."

References
http://epa.gov/biotech_rule/pubs/fra/fra004.htm
HSE 2009. The SACGM Compendium of guidance (Part 2)