Synthetic biology is about more than just slaving away in labs, mixing together tiny amounts of colourless liquids. In particular, our project lends itself well to making an impact on the wider public. Addressing such an important issue as the global energy crisis is a cause worthy of public engagement, which we indeed strived for over the course of the project. It turns out that countless hours spent talking to parents of students at school, writing articles for the school newspaper and presenting talks about our project were not wasted. Mr Zivanic’s Twitter prowess and Jake’s endlessly enthusiastic rambling about our project did not go unnoticed, and by the time Jamboree came around students, teachers and parents alike were engaging with members of our team all the time, asking probing questions about where our money was going as well as raising some of the more serious ethical and moral issues surrounding synthetic biology. Science is meant to be shared with the public and made accessible to everyone, and we believe we have succeeded in this with the human practices side of our project.
In any successful scientific endeavour, safety is of paramount importance. In order to keep ourselves, our fellow humans, and our environment safe strict biosafety regulations were in place throughout our project. However, from the very start there were two aspects of our project that put us in a unique position in terms of safety considerations. As we wanted to do all the work in our school labs, we had to notify the Health and Safety Executive of our contained use of GMOs. This whole process is set up without High Schools in mind, so the challenge of appointing and training the Biosafety Officer, finding the suitable committee to check on all the policies and finally obtaining the permission to start working was something we didn't initially anticipate. Secondly, given that we didn't have the facilities to store chemically competent cells in school, we had to carry out our transformations at Imperial College. This meant that we had to consider the rules and regulations surrounding safe transport of material across London.
Health and Safety Executive
As our PI had the necessary training he could act as the Biosafety Officer within the school. He scrutinised all the risk assessments and finally signed them off. Having done this, we approached the Imperial College team and asked their supervisors if they would act as out GMO safety consultants in order to obtain the permission to start the work. Having addressed their suggestions, we sent off our notification and were allowed to start working.
Transport of materials
Our organisms and DNA were kept securely in the school and Imperial College laboratories and transfer of any biological materials was performed under strictly controlled conditions as per the regulations - double sealed containers to prevent against a leak to the surrounding environment, clearly labelled and protected. Containers were clearly labelled with the details of the two labs between which they were transported and only our PI was allowed to transport these from one place to the other.
In the school labs very few potentially hazardous chemical substances were used during the project and safer supplements for a wide range of processes were used to ensure our safety. With all processes (transformations, ligations etc.) proper sterile technique was used and all wastes, spills and used equipment was instantaneously disposed in Virkon and autoclaved to prevent contamination. All procedures and lab processes were taught to us by the personnel trained in these techniques, after which we were performing them on our own. All the wet lab sessions were supervised by our PI. All the team members were able to effectively carry out safety protocols laid down by the biosafety officer and the handbook. The team was properly briefed on emergency procedures such as fire drills and burn treatments. The entire team took part in a scheduled (and an unscheduled) fire drill during lab time over the summer.
The organisms used in the project were: E.coli DH5α (K-12 derivative) and Synechocystis PCC6803. None of the aforementioned were classed above a risk group 1 by biological safety associations including the ABSA (American Biological Safety Association). None of these organisms, each used in varying procedures, pose any threat to healthy adults. We took care to ensure that none of the chassis used in the procedures were pathogenic and that each was extremely harmless.
Safe project design
Currently, one of the limiting steps is the rate of growth of cyanobacterias the rate of Carbon Dioxide transport into the carbxxysome. By overexpressing the bicarbonate transporter CmpA and linking it to a strong constitutive promoter we are hoping to increase the growth rate of cyanobacteria making it possible for them to be used in shorter time periods. This inevitably poses a potential threat. Should this organism wriggle its way to freedom and find itself in the wild, its increased growth would outcompete other rival species. However, this has been taken into account. By using a replicative rather than an integrative plasmid, we are able to adequately test any increase in BPV efficiency whilst still ensuring that in the worst case scenario (that is, release into the wild) the plasmid will be lost after around two generations and the organism will no longer present any threat to the surrounding environment. More importantly, any equipment used and any organisms after the work had been finished were discarded in Virkon and autoclaved.
Safety when scaling-up
In the event that this were to become a feasible large scale operation it would require an integrative plasmid coding for an increase in cmpA, this would require a control mechanism to be installed with it to ensure that in the event of a release into the wild it could still be controlled. Viable control mechanisms include: Logic gates controlling transcription - only in the presence of specific molecules would the desired gene be transcribed thus allowing us to up regulate the gene. A ‘deadman’ switch, where an external chemical prevents transcription of genes coding for toxins that would otherwise destroy the cell. A CRISPRR kill switch would be viable as well in the event of large scale use of these BPV’s, allowing DNA to be accurately and precisely deleted in the presence of a new environment, having been shown to reduce viable cells by 10^8 fold in the presence of a control substance, this type of kill witch could prove valuable for future large scale work. Ingenious innovative means of control could one day ensure higher levels of safety for an industrial operation, introducing synthetic amino acids into the organism meaning that once the organism is dependent on these synthesised amino acids it would be unable to survive in the wild allowing safe lab use.