QGEM 2022 hopes that our project can inspire and contribute to the success of future iGEM teams. With the development of a prototype, the re-engineering of a polymerase and our entrepreneurial ventures, we hope to act as a steppingstone for future iGEM teams to build off our work.

Our most significant contribution was the successful construction of a physical prototype. The main “kettle” body was welded together at our Universities CNC Machine shop from aluminum. A professional at the shop did the welding for safety reasons, thus we recommend outsourcing to a local CNC machine shop if you require a small, welded part. The lids of our device were 3D printed, however we ran into some issues with the sizing of different parts. We have documented all of our final 3D printed parts for each of the filter, LAMP and drinking components here (insert link to page about different lids). We have also included the ARDUNIO code used for our temperature sensor to monitor the LAMP reaction. We have validated that our temperature sensor works, thus we believe this code would be valuable to future teams who are looking to incorporate a temperature sensor in their device as well.

Figure 1. Physical prototype of our working device.

Figure 2. Temperature sensor code developed for ARDUNIO microcontroller.

It is known that Loop Mediated Isothermal Amplification (LAMP) is a recent emerging technique in the realm of Nucleic-acid Amplification methods. There is tremendous potential for this technique due to its speed of amplification, high yield of amplicon and no need for a thermocycler. Drawing inspiration from iGEM Fudan 2021, we decided to develop our own version of Bst to enhance the LAMP reaction. Through literature, we discovered a DNA-binding protien (DNA-BP) Sac7e. This protien is a homologue to a commonly used DNA-BP called Sso7d which was used in conventional PCR to enhance Taq polymerase. However, it was proved that Sac7e could detect DNA at lower concentrations and is more thermally stable than Sso7d which prompted us to use this DNA-BP. Through the development of our re-engineered Bst polymerase, we were able to theoretically make the enzyme more processive and more thermostable. With this, we hope that future iGEM teams will be able to use, continue to engineer our polymerase or make use of DNA-BP Sac7e if such DNA-BP is required.

Figure 3. Engineered Bst polymerase for increased thermostability and processivity to enhance LAMP reaction, and DNA-BP Sac7e

We were able to successfully express and purify the first iteration of our protein and were able to express our final version in BL21 E. coli cells. With our protocol, we hope that future iGEM teams could optimize the final purification steps in order to successfully purify our final version of Bst.

Throughout the course of our project, we realized how important entrepreneurship and project implementation was to the iGEM competition in terms of creating a project that could actually be implemented and used by target consumers. We created an educational resource brochure outlining important points on financial literacy such that future iGEM teams or even small science/engineering based businesses could reference our brochure to aid in their entrepreneurial journey. To learn more, check out our Entrepreneurship page.