SafetyWould any of your project ideas raise safety issues in terms of:
Our project- ''In vivo Drug Factory'' – involves the use of basic and conventional techniques of Molecular Biology and Genetic Engineering i.e. Transformation, Plasmid isolation, Gel Elution, Restriction Digestion and ligation. Ergo, there are no special risks that the researchers will be exposed to.
Our experiments have been performed in complete isolation and care has been taken to ensure proper sterilization of petri plates, micropipette tips, flasks etc. before usage. Once the work is done, vessels containing cultures were decontaminated thoroughly before reuse. This is ensure safety for the public and other researchers also.
To prevent any harm to the environment, we have ensured that all our unwanted and/or used materials have been disposed of properly. This includes ensuring that the wastes were disposed of in the appropriate Biotech Waste Disposal Truck at the end of everyday’s work.
The following pictures give a few highlights of our safety regime :-
- Plates are properly sealed to avoid cross-contamination
- Isolated Laminar Flow
- Harmful chemicals such as EtBr are properly sealed and handled
Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?
As you would have gathered from out project description, our organism’s presence and role is in the human alimentary canal, particularly the small intestine. There is a particular toxin called enterotoxin which has a particularly marked effect upon the gastrointestinal tract, causing vomiting, diarrhoea, and abdominal pain. So, we have compared all the sequences of our various biobricks to the sequence of enterotoxin (using NCBI-BLAST) to check for similarity on any level. The results are as shown:-
BLAST Results for New Parts
BLAST Results for Existing Parts
As you can see, certain parts showed one or more hits. To nullify this effect we have introduced a kill switch :-
a toxin which is induced by an external molecule – in case of any emergency, the external molecule can be ingested which activates the toxin and thereby kills the e.coli containing the plasmid
use of antibiotics to kill the e.coli cells. Taking into account the damage caused to the native e.coli population in the gut, the use of antibiotics must be followed by an appropriate dose of a suitable probiotic.
Is there a local biosafety group, committee, or review board at your institution?
Unfortunately there is no such biosafety group. So we’ve decided , after due consultation with our faculty advisors, to follow the following biosafety guidelines.
Laboratory Biosafety Manual
Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering? We find that the BioBrick standard could be made much more safer by incorporating the following features in all BioBrick Plasmid Backbones
1)Introduction of our newly introduced hok/sok (ParB Locus) part (BBa_K513000) or other such plasmid partitioning systems, in all plasmid backbones (both for Assembly and Expression). Such a system would improve both the efficiency of BioBrick based systems and prevent the survival of incompletely transformed cells
2)Incorporation of a toxin based kill switch (such as our part BBa_K513001) to prevent the survival of potentially pathogenic transformants.