Team:CU-Boulder/Safety
The main concern in the field of synthetic biology is that the synthesized plasmid will somehow be introduced to the wild and cause toxic or unprecedented effects. It is therefore the responsibility of the designers to ensure a safe design of the plasmids in the lab as well understand the possible scenarios of the plasmid existing outside of the lab. All plasmids that we used are designed to be selected by a corresponding antibiotic. Thus, only cultures grown with their antibiotic will be selected to grow and carry on the formation and incorporation of the protein into daughter cells. Plasmids grown in the wrong antibiotic or with no antibiotic will not be selected for over time. These cultures would only produce DH5 alpha cells with no incorporation of the proteins we introduced; thus none of the DNA we modified would be viable to grow outside of the lab. DH5 alpha cells are a categorized as Biosafety Level 1 and pose no threat to the public. This is an example of how we designed our plasmids with a safe design by introducing the promoter sequence to act as a control.
To image the endogenous Eut S compartments, we grew cultures of the protein and plated these on minimal media plates, which contain the toxic chemical ethanolamine. To control this reaction, we employed the use of the amino acid thiamine, commonly known as vitamin B1, which has been proved to be essential for the DH5 alpha cells to grow. Thus, with no thiamine available, the cells couldn’t not grow or incorporate the toxic chemical into them. In a thiamine free environment, the cells would grow as normal DH5 alpha cells which, once again, are known to be non- pathogenic.
Our lab is classified as a Level 1 in biosafety due to the nature of the DH 5 alpha strain of E. coli cells we use. Non- pathogenic E. coli cells are not associated with disease in healthy adult humans and thus require the minimum safety precautions. Therefore, our lab work for the summer was required to adhere to the rules of a biosafety 1 group including the use of gloves and masks and safe decontamination of waste. In compliance with CU Boulder Environmental Health and Safety, all team members were certified in Basic Biosafety Training, Hazardous Waste Training, Recombinant or Synthetic Nucleic Acid Molecule Training. Furthermore, all procedures as well as DNA orders were overseen by our P.I. Dr. Brain DeDecker.
In accordance to iGEM and federal regulations and laws, all DNA was shipped in a clearly marked container and DNA was packaged in a leak proof primary receptacle within a secondary leak- proof receptacle. All DNA shipped with dry ice contained the words “DRY ICE” on the label and was clearly marked with the appropriate temperature to be maintained at as well as how many kilograms dry ice was used. No shipping papers or documentation were necessary for any contents that we were shipping.
