The first bio bricks made by the NRPUEA iGEM team are hybrid promoters which are composed of a mammalian promoter fused to a bacteria promoter. This has been done in both orientations. Since this is a new concept they are not well characterized, although both individual promoters are well characterized and understood individually.
The first bio bricks made by the NRPUEA iGEM team are hybrid promoters which are composed of a mammalian promoter fused to a bacteria promoter. This has been done in both orientations. Since this is a new concept they are not well characterized, although both individual promoters are well characterized and understood individually.
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Please reference the biosafety level of parts. If you are working with anything other than a BSL1 organism, take extra care with this question. Your nation regulates handling and transfer of pathogens and parts associated with pathogenicity. For a list of regulated organisms, see the Australia Group website.
Would any of your project ideas raise safety issues in terms of:
When designing the experiments the NRPUEA team would undertake we had to consider not only the safety of the researchers and others within the lab, but also the public and the environment.We had to consider carefully the organisms and chemicals we would be using within lab, how we would handle these safely and then dispose of them after to minimize effect on environment and public.
Researcher safety
The university has safety standards which have to be met, which included having basic safety training before starting wet lab work, as well as reading, understanding and signing COSH forms.Therefore, when designing experiments we considered the risks associated and checked whether the COSH forms signed covered the procedures undertaken.The team also received training from advisors over the first few days. They learnt what to do in emergencies, where we could access the safety manual which is always within the lab, as well as being shown where to dispose of the different chemicals and used equipment. It is also vital that researchers within the university ware personal protective equipment (PPE), including lab coat, gloves and covered shoes. The members also wore protective face shields when using artificial UV light in the darkroom. The team were prohibited from eating, drinking or smoking near the lab. The team enjoyed their lunch well away from the lab, as well as being careful to wash their hands before eating and leaving.
The lab facilities were designed to be safe for the team members, providing plenty of space and clean surfaces to work on. The lab was supplied with plastic handheld pipits, and perminant pens; removing the need to use mouth during experiment eg: mouth piptets and no need to lick labels.
The team had originally thought it would be interesting to look at NO levels within salmonella, but decided later that this would result in a lot more safety concerns and complications. Therefore , the team decided to only use strains of "E.coli" (which they had all used within their university studies before and allowed to be handled within the our lab) within our projects. The organism used as a chassis, within NRPUEA iGEM teams experiments, was E.coli ( NEB 5-alpha E.coli was use to characterize existing bio bricks, as well as BL21 pLysS cells and Alpha select gold E.coli. E.coli is a well-studied type of bacteria and therefore has a fairly predictable behavior as well understood pathways. This reduces the risks of danger to the team. E.coli is a non-pathogenic bacteria which has been categorized as a biosafty level 1 bacteria. Therefore, all the students were able to use the E.coli within the university’s second year (category one and two) teaching labs.
Since the team’s project involves Nitric Oxide (NO) they had to consider the safety of the chemicals being used.It is important to note that all the chemicals within the lab are supplied with a safety advice sheet that has been generated by the company. NO has been classed as a dangerous gas which can be directly toxic to blood, lungs, pancreas and nervous system. Therefore, Cautions must be taken when using NO within the lab. It is important to use safely due to its clear appearance reducing the indication of the presence of the gas, possibly allowing the gas to be unintentionally inhaled (suggesting a possible application of an NO sensor for Health surveillance; therefore placing sensor on wall of lab could allow lab to be evacuated if NO present within room reached a certain level). However, the gas has an irritating odor allowing the indication of its presence. Therefore, NO is handled within the lab fume cupboards and chambers are used. This is to reduce exposure to the researcher. In reality, although the team would aim to eventually test the comparator circuit using NO, they will be testing it with promoters which respond to safer and simpler alternatives, such as arabinose sensitive. This allows the team to test the science of the comparator circuit , as well as getting use the procedures before using more toxic chemicals.
When planning the project and the experiments the team discussed and considered their choices of equipment to use. They considered the equipment’s safety to the researcher, as well as the practicality and quality of results gained. Most of the equipment and procedures used within the team’s iGEM project have been previously used, by the team members during the first two years of our undergraduate degree. However, there were a few procedures such as the nanodrop and fluorometer which had not been used before. Therefore the members of the team who were using these pieces of equipment received training from experienced members of staff, in order to avoid accidents which could damage the researcher or the machine itself. Finally, there were always more experienced scientists present within the lab, such as our advisors, who the team could ask for advice on how to use equipment correctly.
Public safety
The team remove their lab coat and gloves, as well as wash their hands before leaving the lab. This is to reduce the chance of bringing anything out of the lab and in to the rest of the university. The team also keep the windows shut, in order to reduce exposure to the outside world.
All contaminated waste is autoclaved on site before being collected and disposed of under controlled conditions for cleaning for reuse.
For the public safety, it is important that we give them the opportunity to understand and ask about what we are doing within the lab, as well as making the advantages and disadvantages clear. The team took the opportunity at the future of science to explain the procedures, such as wearing lab coats that are undertaken.
When the team took transformed E.coli samples to the forum event, we has to consider the public safety. Therefore, we had the plates in a UV box behind a sheet of plastic so that the public did not such them. the plates were also sealed up. Finally, they were kept on a different stand well away form any food ect.
Environmental safety
The materials data safety sheets that come with chemicals explain how to dispose of the chemicals correctly. This is also important as Anglia water carry out random samples on what is poured down the sink drains, and if a certain volume (volume is unknown to university, which encourages them not to put any waste down the sink) of listed chemicals are found, then the university is fined.
consider risks to team members, publics and environment if the project does not go according to plan?
What are risks if safety measures such as containment procedures go wrong and organisms or parts are released? What are risks to security from malicious misuse? How are you addressing such risks?
Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? If yes,
did you document these issues in the Registry?
how did you manage to handle the safety issue?
How could other teams learn from your experience?
The first bio bricks made by the NRPUEA iGEM team are hybrid promoters which are composed of a mammalian promoter fused to a bacteria promoter. This has been done in both orientations. Since this is a new concept they are not well characterized, although both individual promoters are well characterized and understood individually.
Please reference the biosafety level of parts. If you are working with anything other than a BSL1 organism, take extra care with this question. Your nation regulates handling and transfer of pathogens and parts associated with pathogenicity. For a list of regulated organisms, see the Australia Group website.
Did you document these issues in the Registry?
How did you manage to handle the safety issue?
How could other teams learn from your experience?
Is there a local biosafety group, committee, or review board at your institution?
If yes, what does your local biosafety group think about your project?
If no, which specific biosafety rules or guidelines do you have to consider in your country?
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?
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