On this page we discribe about how we are addressing any safety issues in our project.
Safe Project Design
We have set up many safety standards so that, during lab work no organism or DNA exit the premises. One way this is done is through autoclaving the trash before disposal.
Our aim is to produce an enhanced protein. A potential environmental risk comes from the cyclic TALE, which only binds to the target dsDNA. The application depends on the user, for example, a TALE with a nuclease. This TALEN cuts the predicted sequence and should not be taken outside the lab. Therefore our project aims to a much higher safety level than it is possible nowadays.
Safe Lab Work
Before the lab work started, every member of the team got a safety instruction from one of our advisors, Dr. Thomas Reinard. This included general instructions on how to work and behave in a lab, as well as rules to keep the lab clean (lab service). Additionally, he pointed out the location of safety-related objects (such as eye showers, first aid kit etc.), instructions on how to use lab equipment (such as centrifuges, heaters, fume hoods etc.) and, how to properly handle dangerous substances. Lastly, we were taught the record requirement for GMOs, and given a list of rules.
For example we are not to be left working unattended , that is to say at least one advisor has to be present. We must inform someone if we are going to be unusually late and lastly we the possible dangers for expectant mothers were explained to us.
Our team is comprised of students from different study paths, so only those members who were already trained worked in the lab. To protect ourselves and avoid contamination, we always wear safety goggles, gloves and a closed lab coat and shoes.
Our lab has been ranked level 1 in biosafety. The organisms we use are the Escherichia Coli strains TOP10, DH5α, BL21 (DE3) and Origami B (DE3). They are all classified in Risk Group 1 in biosafety guidelines. The vector we are using is the same as the shipment, pSB1C3 .If we work with these strains or use liquid nitrogen to store our samples, we are always using thermo-insulating gloves, Dewar vessels and glasses to prevent any risks.
The gels for the polyacrylamide gel electrophoresis are prepared and run on a separate working place. After this, we dispose of our old gloves and get new ones. Working with genetically modified organisms always contains a risk and, although our experiments are not dangerous, we always work in accordance with the biosafety guidelines of our institution and the government:
It is an ordinance with the security levels and safety measures set for genetic engineering operations by the genetic engineering facilities (Genetic Engineering Safety Ordinance - GenTSV). The ordinance also describes how a disposal of GMOs has to be done.
states the requirements in terms of safety, occupational health, hygiene and work science with respect to activities involving the handling of biological agents. They are drawn up by the “Ausschuss für Biologische Arbeitsstoffe” (ABAS) - which can be translated to Committee for Biological Agents - and adopted by the Committee in accordance with developments.
In Germany, genetic engineering is a topic in the in the Genetic Engineering Act. It concerns all materials who are potential risks. Essentially, it regulates activities of development and application of genetically modified organisms, in laboratories for example, and also governs deliberate releases of genetically modified organisms into the environment.
§1 loosely translated: (1) The 'Biostoffverordnung' is applied every work concerning biological material. It regulates actions to protect the safety and health of employees. Furthermore, it regulates actions to protect people who are not directly affected by the biologic material. (2) The BioStoffV is also valid for work being subject to the Gentechnikrecht (genetic technology law) if there are no equal or stronger regulations on protection of the employees.
5. Allgemeine Stellungnahmen der ZKBS (general recommendations of the ZKBS -Central Committee on Biological Safety)
Our software can produce custom-built TALEs by using Golden Gate Cloning. Since everybody can use different parts, we cannot assure that all custom TALEs are safe. However, they are proteins which will not a risk to the environment if treated carefully. They should not be taken outside the lab.
We did not encounter any unusual safety problems in sending our DNA parts. However, we did follow the protocol provided by iGEM to make sure that there was no risks of contamination.