Safe Project Design
Just shown in the project design chassis section, we chose a safe chassis of E. coli Nissle 1917 and hope to use the genome-modified strain of Nissle SYNB1618 in the future. This strain lacks colonization ability and will be emptied within 48 hours. Experimentally, we only used the common DH5α and BL21 Escherichia coli strain to test our system.
The interleukin-10 which we choose to produce is a natural immunosuppressive agent that is secreted
under controlled conditions, therefore does not cause harm. IL-10 has been used by the past iGEM teams.
For myrosinase, it is a natural component in cruciferous plants. It converts glucosinolates into
sulforaphane to exert anti-cancer and inflammation-inhibiting effects. Similarly, people will be
exposed to sulforaphane when eating vegetables such as broccoli and kale.
Meanwhile, all of our experiments are carried out cautiously, and we don't arbitrarily manipulate model until we are sure that our parts will perform the correct function. For example, we would first use GFP and blue chromoprotein (amilCP) rather than a real-world anti-inflammatory or anti-tumor molecular to establish the model, also we would first use fluorescent proteins indicating our responses/outputs rather than IL-10 or myrosinase.
There are a lot of fears in the general public's mind when it comes to genetics and genetically modified organisms. One problem that arises with most of these systems are not tackling the problem of biorthogonal DNA escaping into the environment after cell death. In view of these security issues, we added a kill switch device to our project. We use cold shock expression of the toxic protein mazF to prevent the harm caused by potentially leaking engineering bacteria.
Safe Lab Work
All the Labs we used contain the correct collective protective equipment. Before we carried out our experiments, all of our team members had received lab safety training, including standard experimental protocols, proper disposal of biological and chemical waste and so on. Each of us, at the moment we got into the Lab, attended to the use of the right individual safety equipment and clothing. Furthermore, in order to ensure complete protection, one has to pay attention to personal hygiene, hand washing is required after each completed job and before leaving the lab.