Team:HUST-China/Safety

Safety

Safety has always been significant to a practical project especially in synthetic biology. This year we generated an auto-consolidation kit mainly applied to marine environment. It brings a huge challenge whether to the strain or to specific environment. However we have shouldered our responsibility to assure the safety of our project and other process in the competition.

Safe Project Design

We deal with the problem by introducing safe strain and safe gene product. All the proteins expressed in the kit have no toxicity to marine organisms and will cause no known diseases to human, plants or animals.

The strain Yarrowia Lipolytica JMY1212 we used as chassis is generally regarded as safe microorganism in safety group 1. It naturally exists in the ocean and intestine of healthy marine fishes. So the compatibility of the strain and marine environment can be reasonably high. The Y.L is also widely used as bioreactor in Industries of Foods and Pharmaceuticals. Based on the previous utilization, normally the genetic engineered Y.L strain still performs stable function because the backbones of plasmid employed was also from Y.L strain.

In flocculating system, our main working agent Mcfp-3 protein is a natural product from Mussels (Mytilus californianus) as component of its filopodia. It enhances the filopodia’s sticking ability to bind strongly to rough surface of rocks or ship-body. This coherent substance shows excellent adhesion performance under water. This property is a double-edged sword, on the one hand it is a strong adhesive material, on the other hand, it may lead to biological contamination considering the Mussels are believed to be fouling organisms. We had thought about this before, and just to emphasize, if designed well, the product of a fouling organisms can be safe and efficient. So we take the advantage of it. By simulating the Euk.cement system with a diffusion model, we put forward a reasonable implementation methods: The engineered yeast was stored in container with a sustained light lamp (inhibition device) . It can be transported to appointed place in a normal concrete truck in which sands and Euk.cement can be mixed in darkness and cementation is started in advance. Or we can also transport Euk.cement to appointed place with light, and then mixed with local sands in darkness to initiate system working. In either way, after about 60 hours mix in darkness, the container together with sands and working Euk.cement can be sink deep underwater to the target region that is sealed with fense. Cementation will finally be performed silently. Besides of fences, the Si-tag displayed on cell surface can further limit the diffusion of cells. So we can put the cementation under control.

As last step of the cementation, CO2 released from respiration of Euk.cement cells will capture abundant Ca2+ or Mg2+ in marine environment and form carbonate sedimentation, the yeast will gradually turn to auto-termination owing to limited respiration and lack of nutrition.

Therefore our kit can reasonably bear the responsibility to preserve the ecological environment and fix the broken basement of the seabed. And with aggregation and growth of marine organisms in the artificial reefs we built by Euk.cement, the ecological environment will finally be full of vigour.

Safe Lab Work

Safety in Lab work is also impartant. Our team has an effective security guarantee on this issue. Before new members enter lab, we organized safety training courses taught by previous team members, graduate students and Principle Investigators in our collaborating lab. Members will be admitted to work in the lab only if they have passed the safety examine. And undergraduate are not allowed to some Level 2 equipment unless they are accompanied with graduate students. We have trained them about the standard experiment operation, safety regulations of equipment use and the good practice rules in the polluted area. Fortunately the strains we employed in our project are all safety Level 1 organism, so most experiments are at Biosafety Level 1. These are several points we emphasize on in our lab safety courses based on the previous experience:
1.Polluted area must be isolated from the normal operating platform. Experimenter are required to wear lab coat and polyvinyl chloride plastic gloves when enter it.
2. Toxic reagent must be recycled specially.
3. Volatile chemicals involving experiment must be operated in the fuming cupboard.
4.  Laboratory appliance must be sterilized before and after use.
5. Safety and maintaining of complex equipments must be taken charged by certain staff.

Safe Shipment

We value the safety in sending parts to registry a very important aspect in competition project. We will never send liquid sample but only frozen dried material will be delivered. The sealed 96 well plate was wrapped with enough bubble paper to make sure it will not be broken. Sometimes the DNA delivery will be checked by customs and certain safety certification may be required to make sure the containing material is not harmful to destination. We obey these rules and prepared materials, such as safety certification, and package according to courier’s requirement. In most delivery, the customs will finally discharge it after checking. This procedure may take additional days and we will take the delay into consideration before sending out our delivery.

In general, yes we understand that a synthetic biology project in real world demands long-term and more comprehensive tests, while here we have only few months to verify the safety of our project. But we can say that according to all data and modeling analysis results we’ve got so far, our Euk.cement kit can be safe to human andenvironment. Of course there is still a long way for the safety issue of our project to go, we are planning to make further efforts to improve the kit’s properties, make it fit better to complex environment.