Team:EPF Lausanne/Safety

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Taxi.Coli: Smart Drug Delivery iGEM EPFL

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Use this page to answer the questions on the safety page.


Risk assessment and safety: TAXI.COLI: smart drug delivery

Overview:

As you might know iGEM is not only about working in a lab, pipetting in lab coats and doing cool stuff with bacteria. iGEM is also about asking yourself the good questions when it comes to the implications and impacts that our project could have on our peers and in our environment. Innovation is not only about having crafty ideas it’s also about opening our mind to the impact of our work and ideas on our environment. We decided that evaluating our project with a risk assessment was a good way of bringing into focus what could be the impacts of our project on co-workers, peers and on nature.

Our project consists mainly on engineering E.Coli in order to transport nanoparticles filled with drug to a specific site of action where the particles would be digested by a release of gelatinase. Thus allowing the drug to enter this particular site where an action is required. An application of such a device could be the treatment of the colon cancer. E.coli being already a part of our gut flora, we thought it could be a good chassis to begin with.

1: Do the biological materials used in your lab work pose any of the following risks? Please describe

a. Risks to the safety and health of team members or others working in the lab? The biological materials we’re working with in the lab are especially thought to be the safer possible for all the team and co-workers. Indeed our plan since the beginning of the iGEM project is to build a project, which couldn’t raise any particular safety issues for us the students but also for our co-workers in the lab. We are particularly interested in designing a project that relies on simple ideas but which is still innovative, original and challenging. Besides our team followed some specific biosafety lectures given by Stéphane Karlen our school safety coordinator. We were particularly careful on wearing permanently in the lab our lab coats and gloves. Indeed working with RG1 biological material doesn’t mean that no precautions should be taken.

b. Risks to the safety and health of the general public, if released by design or by accident? Risk considerations are different regarding what part of the population could be affected. For example in our labs the populations is mainly young and do not have daily contact with young kids, babies or elder. The general public is not the same population than in a lab. This is why we took special care in leaving our lab coats in the lab, not touching the doorknob with gloves and always closing the doors. Furthermore the lab is a confined environment and it is organized in a way that no biological material could get out. Let’s imagine some biological material could still get out of the lab, it couldn’t still cause any arm to the general population since the quantities of bacteria we’re working with are very small and also E.Coli K12 that we are using is not armful nor toxic. Furthermore our labs are restricted to visits.

c. Risks to the environment, if released by design or by accident? The main problem that could exist regarding the environment is resistance to antibiotics. Bacterial conjugation is the horizontal transfer of genetic material by direct cell-to-cell contact or by a bridge-like connection between two cells. This could happen when engineered bacteria containing plasmids resistant to antibiotics are released in the environment thus exchanging their genetic material with wild-type bacteria. Therefore providing antibiotic resistance to bacteria that originally are not resistant. A second potential risk could be the quantity of bacteria released in the environment may disrupt the environmental balance. However we are working with really small quantities of bacteria so the second potential risk is not likely to happen. Still for the antibiotic resistance some precautions are necessary to be undertaken. For example, our entire waist is first inactivated and then burned to prevent any release in the environment by our own waist treatment company in our university. All our used material is autoclaved and sterilized. Furthermore we are working with E.coli DH5alpha (K12), which is a lab strain that wouldn’t survive in the environment.


d. Risks to security through malicious misuse by individuals, groups, or countries? Our labs are under constant control thus preventing any misbehavior or intend to cause arm. Cameras, security agents and co-workers are on constant vigilance. Lab access is restricted at night and after working hours. Our main concern when designing our iGEM project was safety. We wanted to stick to RG1 organisms showing that great things can be accomplished also being respectful of the environment and people. Even in malicious hands our project couldn’t cause arm or cause a risk to the individuals, groups, countries or environment because it is designed to be safe since the beginning of its creation.

2: If your project moved from a small-scale lab study to become widely used as a commercial/industrial product, what new risks might arise? (Consider the different categories of risks that are listed in parts a-d of the previous question). Also, what risks might arise if the knowledge you generate or the methods you develop became widely available? We took into account in the quantitative risk assessment. Meaning that a variation in quantity of the used material shouldn’t make any difference in the biosafety measures we took. Nevertheless if our project moved from a small case study to a commercial product maybe the measures regarding the waist treatment should be even more severe to be sure a high quantity of antibiotic resistance bacteria is not released in the environment. Also when treating a high concentration of bacteria in the lab, the co-workers should be particularly cautious when handling them being careful not to inhale. Although E.Coli K12 is not risky for humans it is always better to be more careful when handling bigger quantities of biological material and even more cautious regarding the waist treatment. Althought quantity shouldn’t have an impact on proper behavior in the lab. Working with small quantities of RG1 in BSL1 labs doesn’t mean one shouldn’t have an appropriate, cautious and respectful behavior towards co-workers, environment and general public.


3: Does your project include any design features to address safety risks? Our project at the moment doesn’t contain any feature to address safety as we are working on a proof of principle project. However we suggest several possibilities to address safety risks considering these two different cases: -Assuming our project could become widely used as a commercial medical application -In case of accidental dissemination in the environment As a design feature to prevent any arm our project could cause in dissemination we imagined engineering our chassis with an artificial amino acid thus preventing it from surviving out of an environment that is not containing the artificial amino acid. In case:

a) Our project becomes a medical application: the patient ingests pills containing an artificial amino acid during the whole treatment with the engineered E.coli. When the treatment ends the patient stops ingesting the artificial amino acid thus not allowing the engineered E.Coli to survive in the colon anymore. We can also think of a way to treat, inactivate or burn the biological waste. The better way would still be to confine the patient during the whole treatment in the hospital so the waist could be directly treated by the hospital itself.

b) Our engineered E.coli are disseminated by accident in the environment: The chassis we used for our project as a proof of principle chassis is not suitable for medical application or to survive in the environment since it is a lab strain named DH5alpha of E.Coli K12. This strain is not able to survive neither in the environment nor in the human body since it is a lab strain. Furthermore the same mechanism as before can be also applied to the environment. Let’s assume our artificial amino acid engineered E.Coli is disseminated in the environment. The bacteria won’t survive considering the artificial amino acids are not present in the environment. The engineered E.coli would then die after a couple of days.

Since this is only assumption we made and a suggestion concerning the feature addressing safety risks it would still be very important to have in mind the safety assessment we explored above and the safety precautions we presented. Though it would be interesting to test maybe next year for iGEM 2014 competition?

4: What safety training have you received? All students at EPFL received specific lectures on biosafety and proper behavior in the lab. All members of the iGEM team followed these lectures and filled the safety form properly. 5: Under what biosafety provisions will / do you work?

a. Link to our institution biosafety guidelines http://sv-safety.epfl.ch http://sv-safety.epfl.ch/files/content/sites/sv-safety/files/SAF_Rules_Vademecum_[E]-1.pdf

b. Link to our Institutional Biosafety Committee http://securite.epfl.ch/safety-en http://sv-safety.epfl.ch

c. Link to our national biosafety regulations http://www.bafu.admin.ch/publikationen/publikation/01614/index.html?lang=fr http://www.admin.ch/opc/fr/classified-compilation/20100803/index.html http://www.admin.ch/opc/fr/classified-compilation/19994946/index.html d. Biosafety Level rating of our lab according to WHO Biosafety Manual

             BLS 1

e. Risk Group of our chassis organism. RG1

For more information and contact about biosafety and our project in EPFL: Stéphane.Karlen@epfl.ch Sabrina.Leunenberger@epfl.ch Charlotte.Broennimann@epfl.ch

Problem: expliquer projet mieux autour de dissemination changement de chassis pour la medical application Image Structure Anglais Autres info →risk assessment e.coli