Overview
Safety has always been significant in synthetic biology, so as a team participating in iGEM 2016, we should shoulder our responsibility to assure the safety of our project and other process in the competition.
Safe Project Design
Synthetic biology offers a stage for researchers to create new genetic machines to solve real-world problems. Along the way, iGEMers met many problems about gene expression such as promoter leakage, expression delay and so on. Researchers are putting forward many new tools to solve the problems such as logic circuits, CrispR, Riboswitches and etc. This year, we designed a foundational toolkit to regulate gene expression based on previous researches. It is like we provide a black box to our end-users(synthetic biology researchers) and they can use it to define the input and output as their requirement. Our toolkit can transform the input signal into output with three main advantages:
1. Threshold settings
2. Stable expression
3. Phase transformation
According to a researche from our potential end-users, they all showed a positive attitude to our toolkit, they would try to adopt such a bi-stable or tri-stable switch to regulate their expression systems.
What our users need to do is to add a sensing mechanism forwards our circuit, for instance, in our application part, we employ plac and patp2 as signal sensors to response to the environment. And iLDH and beta-galactosidase as output to degrade lactic acid for balance.
The safety issues highly depends on the certain application. Our regulation system works in the cells just in gene level. It has nothing to do with toxins, viruses and pathogens. And the only consequence it will bring to the host strain is overloading which make the growth of bacteria slow.
To get full appreciation of gene regulation system, our toolkit just step a little forwards. Logic gate circuits offers the principle of multiple regulations, it provides more possibilities and involves the concept of switch. And RNAi, riboswitches, post-translational modification enrich the methods of gene regulations. What’s more, our circuits provide more specific logic gates and regulation methods. People can make use of it to tell their own story. It is easier for fresh iGEMers to get start with and focus on the application but not the genetic network.
Safe Lab Work
Safety in Lab work is also significant. Our team has an effective security guarantee on this issue. Before new members entering the lab, we organized safety training courses taught by seniors, graduate students and Principle Investigators in our collaborating lab. Members will not be admitted to work in the lab unless 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 team members about the standard experiment operation, safety regulations of equipment use and the good practice rules in the polluted area.
Safety With Parts & Organisms:
Fortunately, the strains we employed in our project this year are all safety Level 1 organisms, and all the parts related to gene regulations will not produce potential toxins and pathogens, so most experiments are at Biosafety Level 1.
There are several points we have put emphasize on in our lab safety courses based on the previous experience:
1. Polluted area must be isolated from the normal operating platform. Experimenters are required to wear lab coat and polyvinyl chloride plastic gloves before entering.
2. Toxic reagents must be collected specially.
3. Volatile chemicals involving experiment must be operated in the fuming cupboard.
4. Laboratory appliance must be sterilized before and after use.
5. Maintaining of complex equipment must be in the charge of staff.
Safe Shipment
We pay great attention to the safety in sending parts to registry a vital aspect of competition. We would never send liquid sample but only frozen dried material. The sealed 96 well plate was wrapped with enough bubble paper to make sure that it will not be broken. Sometimes the DNA delivery will be checked by customs, so we prepared certain safety certification to make sure that the containing materials are not harmful. We prepared, packaged and delivered all the stuff according to the courier’s requirements.
In most deliveries in China, the customs may discharge or check our materials. This procedure may take additional days. And we took the delay into consideration before sending out our parts.
