Safety
Safety regulations in Germany
Working with genetically modified organisms (GMO) involves risks, such as the unintentional release of GMOs in non-GM facilities and the horizontal or vertical gene transfer of genetic material. For example, antibiotic resistances, which are used as selection markers in our laboratory, can be passed on. Another risk is the expression of an unintended protein/RNA/polypeptides, the risk of which is unpredictable.
We live in Germany with a strict Genetic Engineering Law ("Gesetz zur Regelung der Gentechnik (GenTG)"), so the general ethic question to ask is if it is to be allowed to modify other organisms for our benefits. We think that ethical issues concerning our project are negligible, because we work with single-celled and well known microorganisms. There is no testing or experimenting with animals in our project.
The strains we are using are all safety level one (S1). Our project deals with engineering organisms to generate dependencies on each other or lysis mechanisms (System 1: self regulating microorganisms), which makes them unable to survive in a non-optimized surrounding.
All our organisms are optimized for the growth in this dependent system. Because of the dependencies there is only a small chance that the modified organisms could survive in the absence of the controlled culture conditions.
Again and again in the EU the discussion about the correct handling with the so-called genome editing comes up. On July 25, 2018, the European Court of Justice ruled on the subject: Yes, that's genetic engineering. This verdict shows that it is time for science to proactively defend its own underlying worldview. In addition, factual arguments based on data and analysis should continue to be the basis of societal decisions.
Lab safety first
We received a safety introduction from the safety manager of the laboratory rooms we use and we work according to the laboratory rules of the institute. The work with genetically modified organisms sometimes leads to ethical discussions, which are negligible for the application in our project. We minimize risks by applying and upholding the German Genetic Engineering Law (Deutsches Gentechnikgesetz, Article 1 G 2121-60-1 of 20 June 1990 I 1080 (GenTG)), which includes working with GMOs in closed GM facilities, hygiene routines such as disinfect and washing hands, separate waste organization and documentation of each genetically modified organism. We use non pathogenic organisms (S1) for our experiments, so we do not risk harming anyone if they were to get in contact with them, which of course should not be the case. Our waste containing the modified microorganisms is stored separately in S1 waste containers and will be sterilized by autoclaving before it gets disposed of.
Our organisms
| Species Name | Strain | Genotype | Risk group | Disease risk | Source |
|---|---|---|---|---|---|
| E. coli | K-12 TOPTEN | F- mcrA Δ(mrr-hsdRMSmcrBC)φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(araleu)7697 galU galK rpsL endA1 nupG P3: Kan R; AmpR; (am) Tet; R (am) | 1 | no | Grant et al. 1990 |
| S. cerevisiae | BY4742 | MATα his3Δ1 leu2Δ0 lys2Δ0 ura3Δ0 | 1 | no | Brachmann et al. 1998 |
| E. coli | DH5α | F– φ80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK–, mK+) phoA supE44 λ– thi-1 gyrA96 relA1 | 1 | no | DSMZ |
| E. coli | Pir | F- ∆lac169 rpoS(Am) robA1 creC510 hsdR514 endA recA1 uidA(∆MluI)::pir-116 | 1 | no | DSMZ |
| E. coli | BL21 | B F– ompT gal dcm lon hsdSB(rB–mB–) [malB+]K-12(λS) | 1 | no | NEB |
| E. coli | BL21(DE3)C43 | F–ompT gal dcm lon hsdS B(rB-mB-) λ(DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5]) | 1 | no | NEB |
| E. coli | BL21(DE3)C43 (-leuB) | F–ompT gal dcm lon hsdS B(rB-mB-) λ(DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5]) (-leuB) | 1 | no | NEB |
| E. coli | BL21(DE3)C43 (-leuB, lysC) | F–ompT gal dcm lon hsdS B(rB-mB-) λ(DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5]) (-leuB, lysC) | 1 | no | NEB |
| Synechococcus elongatus | sp. PCC 7942 | Wild type | 1 | no | Genome.jp |
| Synechococcus elongatus | sp. PCC 7942 | ++invA, glf, IdhA, IIdP | 1 | no | Aem |
| Synechococcus elongatus | sp. PCC 7942 cscB:::NS3 | M744_RS12430::Plac-cscB-Cmr | 1 | no | Aem |
| Zymomonas mobilis | Wild type | 1 | no | DSMZ | |
| Corynebacterium glutamicum | Wild type | 1 | no | DSMZ | |
| Pseudomonas putida | KT2440 | rmo- mod+ | 1 | no | DSMZ |