General Lab Safety
The University of St Andrews iGEM team operated in full compliance with the rules and regulations outlined by the iGEM competition. The wet lab carried out all experiments in the lab spaces of Dr Ulrich Schwarz-Linek and Dr Clarissa Melo Czekster, both of which are located in the Biomolecular Sciences Annex of the University of St Andrews. These labs are biosafety level 2, which was sufficient as all iGEM work was carried out using the Origami, DH5a, and BL21(DE3) lines of Escherichia coli K-12, which is a biosafety level 1 organism1.
Before entering the lab each team member that would be working in the lab attended a safety talk by Dr Magnus Alphey, the safety liaison for the school of biology, and successfully completed an online test detailing fire safety procedures, appropriate use of PPE in the labs, and safe working practice. Once in the lab, team members were required to wear appropriate attire, which included lab coats, goggles, gloves (when appropriate), closed toed shoes, and tying long hair back. All waste was disposed of properly in the appropriate bins, and all cells were treated with detergent or verkon prior to disposal.
Antibody CH3 Domains (Parts BBa_K2929000-2)
As the future use of our product would hopefully involve its use in humans, we considered the possible safety issues that may come from doing so. To do this we researched and wrote up an impact analysis, which took into consideration the known off-target effects of past, current, and currently in development stage antibody therapies.
Thermatoga beta-glucosidase A (Part BBa_K2929003)
Thermatoga beta-glucosidase A is derived from the Thermotoga maritima hyperthermophilic organism and as such possesses no properties that require biosafety precautions other than general laboratory safety procedures. BBa_K2929003 mutants have no intended future uses, and since the expression of possible isopeptide-bond containing mutants was only carried out as a proof of concept, there are no possible safety issues relating to the future use of these mutants.
mOrange + SpyTag (Part BBa_K2929004)
mOrange is a fluorescent protein derived from Discosoma species and possesses no properties that require biosafety precautions. SpyTag is a short peptide sequence (13 amino acids) originally derived from Streptococcus pyogenes, but due to its small size possesses no pathogenic properties and contains no Streptococcus DNA sequences associated with pathogenicity. The SpyTag sequence can be used safely under biosafety level 1 conditions. The intended future use of the mOrange-SpyTag fusion only entails laboratory use as a reporter protein and hence has no future biosafety issues.