Working in the lab means that we have safety guidelines! First of all, lab coat and gloves are mandatory along with safety glasses where needed. We start every experiment cleaning our workbench with ethanol and end that same experiment doing exactly the same, preventing growth of contaminants. Furthermore, we separated the benches used for E. coli and Yeast to avoid contamination. When working near the flames we avoided using gloves. All bacterial and yeast cultures were neutralized with bleach before disposal and put in the biowaste. Posttreatment of the waste was done by the home institute facility and according to the legislation in our country.
We are working with dCas9, a catalytically dead version of CRISPR-Cas9, which unlike its active form isn’t able to cut DNA sequences but it only binds to them depending on the guide RNA (gRNA). Therefore the danger from off-targets is already much lower. That did not stop us from checking for any binding sequences for gRNAs in the yeast and human genome (even though we do not intend to use our gates in humans) to add another layer of security. The risk in this could be unwanted inhibition/activation of some of the genes in our own genome! Luckily, we were able to design and obtain gRNAs with no such human-binding capabilities!