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Team:Freiburg/Safety/safety
From 2013.igem.org
Safety & Security
Declaration of Intent
Hereby we, iGEM Team Freiburg 2013, state, that all the DNA constructs used, as well as our mammalian cell lines, were solely utilized for laboratory work. The cells provided us a well-established model system for scientific research, which is similarly utilized by thousands of other academic institutions around the world. Notably, in none of our experiments we used stable integration of DNA-sequences into our host cell genomes and therefore, have not created genetically modified human cells. We purposely avoided the utilization of viruses as DNA-donors, and instead, were strictly relying on transient PEI-transfections in every approach. As Cas9 - the heart of our toolkit - was also mutated to a non-cutting DNA-binding device from the very start of the uniCAS project, the risk of off-target mutagenesis in our mature HEK-293T, HeLa, CHO-K1 or NIH/3T3 cultures was efficiently reduced to a minimum.
Safety
At the beginning of our project, it was deliberately intended to strictly work on a transient experimental scale. This was due to our intention to firstly test for the possibility to regulate and control endogenous processes and to classify off-target effects. Accordingly, we never stably integrated our dCas9 or RNAimer constructs into a human cell genome and did not have to use randomly integrating retroviruses. In regard to biosecurity concerns, our uniCAS toolkit is exclusively useful and intended to act as tool for laboratory research, so purposes to release uniCAS-transfected cells into the environment could on the one hand be judged senseless but also futile. Furthermore, it is worth mentioning that our project doesn’t involve expression of any toxins or pathogenic proteins - whereby secondary risks could be excluded from our project.
Nevertheless our main protein, dCas9 and also its related RNAs, originated from a pathogenic bacterium (Streptococcus pyogenes). This procaryote is classified, according to German regulations, to be safety level 2. We were solely operating with the protein and RNAs vectors - originally distributed from addgene -, which to date have not been described as a part of any pathogenic or toxic activities of this bacterium, they could be considered as nonhazardous. An up-scale of this system would not raise any additional dangers and could be performed under S1 conditions as our small-scale approaches. This is in accordance to German regulations for biotechnology and safety. For a detailed look on our Safety explanations and official forms, please have a look on our safety page.
General Security
Most of all new technologies and methods bring benefits but also risks with them. Therefore, we asked several Freiburgian experts from different biological and medical fields for their opinion - regarding research, ethics and safety. Especially in the context to our project and in regard to multiple gene regulation in general, we received a broad range of optimist but also critical commentaries.
Exemplarily, we intend to point out some key reflections on the topic, which resulted from an interview with Brain expert Dr. Jochen Holzschuh from the Freiburg Department of Developmental Biology.
“... even when considering the highest risks, it is not an alternative to stop research ...“
“... as a scientist who creates a certain knowledge, responsibility does not stop at the end of his or her own work - instead, the duty of observing the usage of this knowledge by other scientists arises ...“
“… a scientist can never stay objective in regard to his own project. Thus, safety judgements always need to be developed with non-involved experts ...“
There have been different methods for regulation of genes before. Hence, we did not set up a totally new branch of research, which would have implied new safety issues. But nevertheless, we engineered the CRISPR/Cas system, and thereby gave rise to the opportunity of a more complex type of gene regulation - because it is now possible to simultaneously target and therefore regulate multiple genes at once. Still Prof. Dr. Wolfgang Hess confirmes that:
“… considering ethics and biosafety: it’s „just“ another tool to do things that have been done before, expect for the complexity …“
One question in our interviews dealt with the problem which security aspects can be seen in regard to laboratory or environment while using our toolkit. Most of the experts emphasized that we did not use any viruses or stable transductions as described above. For more information about the ethical background of our project and the expert opinions on that controversial topic, especially the medical point of view, visit our ethics page.
