Achievements
During the course of this year’s project, our team has achieved many important milestones, which we further elaborate on throughout our Wiki. This page serves simply as a way to center and summarize our achievements.
Wetlab
Our experimental design began as bold ambition: to fully characterize the Receiver-Promoter cross-talk between six Quorum Sensing systems, through a 2-plasmid system with a range of 12 total plasmids. Cloning problems and DNA synthesis problems made us fall just short of the 12 plasmid mark, but we nevertheless generated significant amounts of data and made solid advances in the characterization of the used parts. Our wetlab achievements were as follow:
- Successfully cloned the pLux Ratiometric Activity Reporter Plasmid
- Successfully cloned the pLas Ratiometric Activity Reporter Plasmid
- Successfully cloned the pRhl Ratiometric Activity Reporter Plasmid
- Successfully cloned the pCin Ratiometric Activity Reporter Plasmid
- Successfully cloned the pTra Ratiometric Activity Reporter Plasmid
- Successfully cloned the pRpa Ratiometric Activity Reporter Plasmid
- Successfully cloned the Lux Synthase-Receiver Plasmid
- Successfully cloned the Rhl Synthase-Receiver Plasmid
- Successfully cloned the Las Synthase-Receiver Plasmid*
- Measured and quantified the cross-interaction between the Plasmids we had, with the help of our mathematical model
*The Las Synthase-Receiver plasmid was cloned, but did not activate any Activity Reporter, including the Las Reporter. We proceeded to test it with direct induction by HSL, and it also did not activate the Reporter, so we infer that the fault lies in the LasR Receptor, either in the part itself or on cloning problems on our end.
Measurement & Improvement
With so much of our project revolving around fine-tuned promoter induction, measurement was a top priority for us from the get-go. And so, we are very proud to have designed, cloned, and validated our YFP-CFP Reporter, improving the use of the pre-existing YFP reporter. We hope that our advances may become a handy and reliable reporter tool for future synthetic biology projects.
Modelling
Using as a proxy the results from our wetlab, we've been able to construct a mathematical model and effectively find values for the crosstalk between the systems! We established a relative parameter, lambda, with which we compare those systems in a more precise way, instead of looking at absolute numbers that don't carry any real meaning. With that in mind, we've been able to create a model with 1 to N systems and run simulations for 2 systems. Furthermore, we proposed a statistical analysis for applying the model to real-world data.
Integrated Human Practices
To go along with our lab achievements, we also achieved many important out-of-lab goals. Mentoring the high school seniors, Dani, La and Elo, was truly a wonderful experience for all of us. We have certainly learned as much about mentoring, teaching, and scientific communication as they have learned about molecular and synthetic biology.
On top of that, bringing the topics of synthetic biology and quorum sensing both to specialists and to the general public allowed us to receive valuable feedback on our project, which was determinant for its final outcome.
Interlab
As we further elaborated on our Interlab page, we have successfully taken part on this year’s Interlab Study, providing validated results and keeping in contact with iGEM HQ the whole way through, to make sure everything was done according to protocol. It’s no secret that we are huge fans of the iGEM Interlab Study, as it takes very important and impactful steps towards a more reproducible science.
Quorum Sensing Collection & Registry Parts
Through the course of our project, our team has designed and added to the Registry a grand total of 40 new BioBricks, both Basic and Composite parts, and have acquired substantial characterization data for them.
Along with our new parts, many Quorum Sensing parts and devices already existed on the Registry, and so we decided to make use of all of our previous research to gather them on one handy place. We have therefore created the Quorum Sensing Collection, gathering the parts we found relevant, in hopes that it can be useful for future teams.