Team:EPF Lausanne/Safety
Why is there a need for safety?
Our project is based on the dCas9, a derived form of the very polemical CRISPR-Cas9 protein which raised safety issues concerning the “off target effects”. (when the Cas9 is guided to and cuts an unwanted sequence in the genome) These effects led us to think about some ways to minimize the risks of unwanted targeting, both in the genome of E.coli and S. cerevisiae we are working with. Even though we won’t introduce Cas9 or dCas9 into a human cell we wanted to check if there was a risk of gene activation/inhibition if we got a large quantity of these proteins into our body. In our case (with dCas9), the risk is the following: if a guide RNA causes dCas9 to bind a regulatory region of gene in an unwanted fashion, it might be deadly for the organism.
What did we do to reduce the risk?
To assess these issues we designed a program that automatically blasts the guide RNA (gRNA) plus de PAM sequence (either CCn at the beginning or nGG at the end) against the genome of E. coli, S. cerevisiae and the human genome. When we had the result the program would give us the sequences that are not in the organisms. Note that we wanted to work with completely synthetic gRNA to have as little interference as possible. Future iGEM teams could use our software to read their BLAST results and to draw conclusions of the results that could maybe result in less “off-target effects”.
Experts point of view
We also asked the experts we interviewed about the safety issues raised by a potential use of the BioLOGIC system outside of the lab. What would be the consequences of using BioLOGIC in pharmaceutical industry? Peter Oliver, the Head HTS group at Actelion Pharmaceuticals Ltd told us that while new gene editing or regulating techniques are very useful in the creation of biological models for diseases such as cancers, utilizing them as therapies is not considered at the moment, as the available technologies are not safe enough and the general public (being currently very critical towards the industry) would not agree with it.
