Gold Integrated

Introduction

Follow up from the action plan, we developed a toolkit for engineering kill switch in an effort to address the issue of environmental and health safety concerns raised by our community from our human practice silver results. Further, we have also developed 2 basic troubleshooting guides (Table 1-3) to improve our genetic circuitry design by following some of the design considerations listed in the guides. This would also address the concern of a lack of standardised methodology in building a kill switch. These guides have helped us to better consider the different aspects of designing a kill-switch, including what if the kill switch fails when the engineered probiotics in the body (a consideration which we didn't think of at the beginning of the project!).

Development of Troubleshooting Guide

Apart from the extensive modelling support our team has incorporated into the methodology, we desire to create a user friendly guide that addresses the fundamental problems in integrating the kill switch mechanism into the microbes. The intent is to enable users to troubleshoot potential problems quickly and efficiently.

The features of the guide are:

Opensource and easily available: after registering relevant credentials, any member in field of synthetic biology is welcome to contribute his/her user-experience in the challenges faced in the design consideration aspect, genetic information to this toolkit
Peer-reviewed: fellow experts in the field of synthetic biology can validate one another’s user experience, provide solutions, all in one place.

To gain more inputs for our guide, we initiated discussions with researchers from the Synthetic biology for Clinical & Technological Innovation (SynCTI) on potential challenges in incorporating kill switches into bacterial host to validate our guide (Table 1 and Table 2). We drafted a second guide that considers the possible failure of our kill switch in and out of the human body. This second guide (Table 3) address the environmental and health consumption concerns that were highlighted from our interactions with members of the public, conservationist and experts in the field of synthetic biology.

Table 1. Making kill switches easier to incorporate into bacteria host(part1)

Table 2. Making kill switches easier to incorporate into bacteria host(part2)

Table 3. Considerations for possible failure of kill switch

We have carefully considered both guides that we have developed while developing our kill switch (i.e. choosing a toxin-antitoxin system) and performing our experiments. These guides have aided us in making a decision on the design. However, we did not manage to characterize all of the possible problems listed in the guide due to a shortage of time. Our team hopes that both guides will serve as a basic guiding tool for future kill switch gene circuit design considerations and for the scientific community to improve on both guides to create an even more extensive guide!

Conclusion

We have developed a methodology comprising 2 troubleshooting guides with a modelling methodology to address the environmental and human health safety dangers that genetically engineered organisms post, as well as the lack of standardized methodology in integrating a kill switch safeguard into bacteria chassis. In terms of future work implementation, we hope to get this entire system online to facilitate an open source, accessible repository to the world in alignment with the iGEM spirit and that our project will be a key milestone in paving the way to ensure a wide spectrum of futuristic applications that are environmental and health friendly.