The Laboratory in which the iGEM Project has been conducted, at the Chair of biological chemistry is classified as a laboratory with the biosafety level S1. According to the German law of the regulation of GM technology (Gesetz zur Regelung der Gentechnik) there are 4 levels of biosafety, namely from S1 to S4. In the Level S1 there is considered that no risk for human health or the environment is present.^[2] In order to guarantee the integrity of all the team members and the environment, we took preventive steps prior to the beginning of our lab work. All team members that were going to be performing wet lab tasks were required to attend to a lecture about the risks and precautions while working with GMOs. It was conducted by Dr. Martin Schlapschy in charge of the safety of the laboratory. In this presentation information about the regulation of biology by governmental institutions in Germany and the obligations each laboratory has to fulfill e.g. having a good and ethical practice was also included. After this all team members was given an introduction to the laboratory, in order to get to know all important safety features as well as localisation of the proper lab equipment. Factors like disposal of chemicals, GMOs and handling of dangerous chemicals or sensitive cells (cell culture) were specifically emphasized and thoroughly explained.
It is important to underline some important safety measures that have to be taken while conducting molecular biology experiments:

• The use of Ethidium bromide, since it is considered to be a mutagenic chemical compound. The use of gloves, and safety googles was obligatory while using it. ^[3]

a-rna-purification- analysis/nucleic-acid-gel-electrophoresis/dna-stains/ etbr.html This measures also applied for compound like Hydrochloric acid and Ammonia, additionally this were to be handled with the protection of a fume hood, due to the volatility of some of them.

• The use of a special protective mask in order to protect the eyes from the UV Light while analyzing gels.
• The use of certain machines at the laboratory posed some risks as well. As an example the use of big centrifuges, which if mishandled could cause harm to the investigators.

In the daily work with genetically modified microorganisms and cells it is always important to consider various aspects of biosafety. When planning and conducting experiments one has to evaluate all potential risks and dangerous posed to human health and the environment thereby and take all eventual necessary precautionary measures (precaution principle). Under this principle general contact with biological material for instance shall be reduced to a minimum. As our project relies on a variety of wet lab work all constructs and experiments were carefully checked ahead of execution. Our work in the laboratory was determined not to pose any kind of unusual risks neither to the experimenters nor the general public or the environment. The strict containment of the experiments within the lab was ensured at all times. All strains and lines organisms and cells used were specifically designed for lab applications and are therefore not able to survive outside laboratory conditions as no aspect of our project work overrides these restraints.

In order to fully ensure that a project does not allow for any harm to be dealt it is also imperative for the scientists to assess possible harmful misuse of an invention. Relevant to such a analysis are whether the genetic construct can be used directly or indirectly to arm microorganisms. Constructs for example that allow for the production of any kind of toxins or dangerous chemicals as well as genetic code that allows for the integration or transmission of DNA.
As we have evaluated our project accordingly we were not able to find any aspect of our work which could be used or modified to serve exterior unwanted purposes. No part allows for the production of any kind of harmful substances nor the spread of the construct themselves.

The topic of bioprinting, due to the fact of being relatively young, is still currently being discussed in international ethics and safety forums and in courts. This is why we consider it to be essential to briefly present and discuss the main concerns around bioprinting. It is important to mention that there is a broad spectrum of perspectives and disciplines, in which key questions about tissue printing can be analyzed.
The world is facing a lot of challenges nowadays with the constant improvement and advances in the field of biology. These being questions about the ethic of some projects and furthermore the regulation of the reproduction of the same experiments, in the case of its success. This is why it is very important to consider all these political, social and ethical issues bonded to the scope and effect of the project. It is worth mentioning how there is a big debate about organ transplantation already since it first became possible in 1945 when the first kidney was successfully transplanted (HRSA). ^[4] With bioprinting a lot of new questions around patentability, affordability towards the different social strata, clinical safety and social acceptance arise and are to be discussed and dealt with.
One of the biggest security concerns around the use of 3D printed organs for transplantation is the tort liability of physicians, added to the intellectual property debate of 3D produced organs. Considering the fact that the transplantation of these organs needs still years of research of becoming a reality, the governments and regulation organisms are facing a great challenge in regulating the use and manufacture of 3D printed organs. To illustrate this problem, as an example, the Food and Drug Administration (FDA) is in charge of regulating medical devices as artificial hearts. Meanwhile is Organ Donation regulated by another governmental agreements, like in the case of Illinois, the Anatomical gift act. ^[5]

Another important factor to consider and discuss is the patentability of such a printer, or in the given case of the printed products. This is where the question comes, whether or not a printed organ should be considered as artificial or normal organs.

One key factor of guaranteeing the safety of our project and preventing in advance any dangers, is the implementation of a well-known kill-switch. In this case we worked in cooperation with the iGEM-team of Slovenia and Freiburg, who each provided us with their Biobrick (BBa_K404113 - HSV-thymidine kinase fused to mouse guanylate kinase, iGEM Freiburg 2010 and BBa_K782063 - CMV-promoter, iGEM Slovenia 2012). This specific kill switch is known as the Thymidine kinase / ganciclovir system. In order to enhance the current Biobrick, we have repaired a point mutation that led otherwise to a pstI restriction location. An additional Poly-A-tail should also stabilize the mRNA later on. For more details, see our experiment protocol further down.

1. ↑ U.S. Department of Health and Human Services (2009, December). Biosafety in Microbiological and Biomedical Laboratories. Retrieved October 6, 2016, from http://www.cdc.gov/biosafety/publications/bmbl5/BMBL.pdf
2. ↑ Bundesministerium der Justiz und für Verbraucherschutz. Gesetz zur Regelung der Gentechnik (Gentechnikgesetz - GenTG)§ 7 Sicherheitsstufen, Sicherheitsmaßnahmen. Retrieved October 06, 2016, from http://www.gesetze-im-internet.de/gentg/__7.html
3. ↑ Ethidium Bromide. Retrieved October 06, 2016, from https://www.thermofisher.com/de/de/home/life-science/dn
4. ↑ Timeline of Historical Events Significant Milestones in Organ Donation and Transplantation. Retrieved September 03, 2016, from http://www.organdonor.gov/legislation/timeline.html
5. ↑ Park, M. H. For a new heart, just click print. Retrieved September 10, 2016, from http://www.globalforumljd.org/cops/legal-and-ethical-aspects-human-organs-3d-bioprinting