Instead of using fish as a chassis, iGEM Peshawar decided to use bacteria in our experiments and give a proof of concept using bacteria as a chassis. iGEM Peshawar used DH5-Alpha strain, which is also known as lab strain. This strain has no pathogenicity. DH5-Alpha is safe to use by human. This strain can be easily controlled under the bio-safety level 1 and 2. This engineered bacterium will not be exposed or tested outside the lab to prevent the horizontal gene transfer to the naturally occurring bacteria.
All the parts that we are using in bacteria are not pathogenic, though the plasmids have antibiotic resistance genes.
In case we make changes in fish in future, we have designed a cage which will prevent the fish from encountering with natural breed (existing species). The case will only allow water to pass through the cage but it will prevent sperms and eggs to pass through it. By controlling permeability of material of cage, we will minimize the chance of transferring of gene to the wild species.
Per international biosafety guidelines our lab comes under the category of Bio-Safety level 2. It is a secured area equipped with an ante–room (buffer zone) and magnetically sealed doors with electronic identification system allowing access only to the approved personnel with biosafety training. Anything that goes out of or comes inside the lab is strictly regulated. No materials can leave the lab without first going through a safety and security check. Our institute enforces WHO biosafety guidelines, more specifically Laboratory Biosafety Manual - Third Edition, as well as our Governmental safety guidelines.
Before starting our wet lab, all our team members had a lab training session. They learned about the bio safety levels and the rules that will be followed in the lab during the project. we received extensive lab safety training from our PI, Dr. Faisal F. Khan, our mentors Muhammad Ismail Bakhshi and Sarah F. Khan.
Throughout our project, we have been following safe lab practices as trained. All work is done while wearing a fully buttoned lab-coat, closed toe shoes and nitrile-coated gloves always. The benches were regularly disinfected with 70 % ethanol. Biological waste was autoclaved first and then it was disposed properly.
Micropipettes, flasks graduated cylinder, gel trays, gel tanks and all equipment that is involved with the ethidium bromide were placed in a separate labelled bench, to prevent any kind of mishaps. All other materials were also clearly labelled with appropriate warnings.
Good microbiological practices were followed; all work with bacterial cultures and plates was done in the Bio–Safety cabinet. U.V light was activated before and after performing experiments in Bio-Safety Cabinet. The incubator was sterilized with 70 % ethanol every month. Plates containing bacterial cultures were Para-filmed and were stored at 4 degrees centigrade.
Special precautions were taken when handling hazardous materials, harmful chemicals and salts. These include concentrated acids and our test heavy metals salts. All chemical reactions were and are being done in the Fume hood with complete protection including goggles.
Our project and all the parts are safe to use. They cannot be misused.