Team:William and Mary/Collaborations
Collaborations
We met the Pittsburgh iGEM team in late July during Mid-Atlantic Meetup and we were fascinated by their project. They are aiming to implement intracellular logic gates using split inteins. It occurred to us that intein splicing could be incorporated into our living materials toolkit as an efficient method of induction if we could control when the splicing event happens. Thus, we reached out to the team for collaboration. During discussion, we were informed that it is possible to incorporate a light-sensitive LOV2 domain into an intein to control its splicing. Upon illumination, the light-sensitive domain will trigger the intein to splice out itself and restore the function of its host protein. We decided to experiment with our LuxR protein by inserting intein DnaE with the LOV domain into it as a part of our engineered quorum sensing system. By controlling LuxR function restoration, we could control the activation of pLux and thus any gene of interest downstream from it. Junction site and intein sequences were designed by the Pittsburgh team, while we were focusing on designing gibson assembly for insertion. We have begun work on testing out the functionality of intein inserted-LuxR.
W&M's project was to repurpose biofilms as living materials. To ensure robustness of such biomaterials, we have constructed a library of biofilm forming and strengthening adhesins for use in E. coli biofilms. This library builds upon AG43, SaSuhB from S. aureus , and Functional Amyloid in Pseudomonas (FAP) from P. aeruginosa . However, we wished to investigate strengthening the adhesion of these proteins even further. We therefore used the tools developed by the St. Andrew's iGEM team to investigate the list of double salt bridge proteins to determine if any of our adhesins were on this list. These bonds are often used in prokaryotic proteins that are secreted and they are known to increase stability and extend protein half-life. This could be an important feature for strengthening and stabilizing biofilms. Although our adhesins were not represented, this information was important for our project because it allowed us to better understand the nature of the biofilm adhesion and design stronger adhesins for our future directions.
As the goal of SmartFilms is to produce a flexible and powerful biomaterials toolbox, we wanted feedback on how it worked in an actual application. Cornell iGEM’s project aims to build a system which is able to detect and filter out microcystin toxins present in water produced by harmful algae blooms (HABs). We saw the opportunity for a strong collaboration in which we gained insight into practical applications for SmartFilms and Cornell would be able to trap more microcystin by using our adhesin library to increase bacterial density.
After reaching out to the Cornell team and receiving an enthusiastic response, we held a conference call to better understand both our projects. We sent Cornell parts for IPTG-inducible curli fibers which were then tested against a control group. Cornell found that the bacteria with induced curli fibers produced 2.5 x 10^6 CFUs as opposed to the control group’s 4 x 10^5 CFUs in a serial dilution, suggesting that curli fibers are able to increase bacterial density, which would allow for more effective water purification.
We also received Cornell’s protocol for entrapping bacteria in alginate beads and we were able to replicate it with bacteria containing pDAWN + AG43. This method of forming alginate beads opens up further applications for our toolbox, as alginate beads can provide a way to transport our biomaterials, or allow for condensed biomaterial factories.
We shared our curriculum with Virginia iGEM for review and collaboration. Considering the updated SOLs affect our entire state, we were eager to involve Virginia teams to collaborate on developing our curriculum. After writing a curriculum draft, we sent it out to Virginia iGEM to be peer reviewed. After receiving detailed feedback, we revised our work based on their critiques. We then sent them our final draft for review, and they agreed that the curriculum was ready to be published. It is freely available on our page. This bidirectional collaboration will improve synthetic biology education in our state by providing teachers across the state with materials to teach students about synthetic biology.
We collaborated with OUC-China by sending them architectural diagrams of our current lab, and our dream lab. By planning out our dream lab, we are actually able to implement our ideas by providing feedback to the architects that are currently designing our new integrated science center, which will house the new bioengineering lab.
We were inspired by Lambert iGEM’s work with the Thirst Project so we decided to help contribute to building a well in Eswatini. Furthermore, we helped provide Lambert iGEM with feedback on their hardware by completing their survey.
Warwick iGEM shared their Biology 101 pamphlet with our team which explains the basics of biology. We reviewed the pamphlet, and thought their original illustrations would be a wonderful addition to our curriculum. Warwick approved the use of their illustrations in our curriculum, greatly enriching our lesson plans.