Bronze Criteria
| Criteria | Description |
|---|---|
| Competition Deliverables | We completed all the competition deliverables (Wiki, Presentation Video, Judging Form, Judging Session). |
| Attributions | We wrote an attribution page in standard form, clearly stating anything that was not done by us students. We clearly acknowledged all the people who helped us with our project on this page. |
| Project Description | We wrote a description and inspiration for our project. |
| Contribution | We contributed to iGEM in many ways: 1. by developing a guide for future teams on how to effectively use the DLS tool 2. by creating the CAPTURE plotting tool, which will allow future iGEM teams to create uniform graphs for their data presentation 3. by producing a collection of informative videos with detailed protocols for liposome preparation methods, LNPs production through extrusion and analyze them with DLS measurement settings (SOP file), so that future iGEM teams can use them. |
Silver Criteria
| Criteria | Description |
|---|---|
| Engineering Success | We went through several iterations of the engineering design cycle for multiple parts of our project, carefully described on the engineering success site for each part of our project. |
| Human Practices | We discussed our/possible project(s) with a bioethicist early on. This led us to the question of who we want to help and what we want to promote. Finally, we chose values as a team that influenced our decision making in economic, social and research interest. Being in the infectious diseases village, with main focus on clinical application, we considered what we would have to do to apply our project to the real-world and summarized our considerations in the treatment implementation. |
Gold Criteria
| Criteria | Description |
|---|---|
| New Composite Part | We did a detailed characterization of our New Composite Part, HSTII-Sushi, and set a foundation for further improvement. We conducted proof of concept experiments showing the functionality of parts of our CAPTURE system. |
| Measurement | We devised a novel approach for rapid and cost-effective quantification of DNA in LNPs. |
| Integrated Human Practices | We engaged with experts, researchers, and healthcare. We used dialogues to face perspectives on certain topics to improve our project’s good scientific integrity and practice. This helped us gain new insights and perspectives that are leading to new considerations regarding CAPTURE. We engaged in interdisciplinary dialogue, aiming to make CAPTURE, with the help from research, society and industry, a more sustainable, fairer and effective product. |
Most Important Achievements
Lipid-based Nanocarriers
- We successfully generated Giant Unilamellar Vesicles (GUVs) encapsulating plasmid DNA and employed extrusion techniques to achieve size reduction.
- We produced and characterized Lipid Nanoparticles (LNPs), verified plasmid encapsulation and quantified it through fluorescence-based assays.
- To test the targeting mechanism, we visualized the interaction between our lipid-based nanocarriers and P. aeruginosa using confocal microscopy.
- We assessed the effects of our lipid-based nanocarriers on A549 lung epithelial cells, and revealed no significant biosafety concerns.
Antimicrobial Peptides
- We successfully showed the functionality of HSTII as an extracellular localization signal.
- We showed the bacteriostatic effect of the expressed HSTII-Sushi in IPTG inducible E. coli.
- We were able to show the effectiveness of the L-enantiomer of the AMP D-CONGA-Q7 for the first time.
OMVs
- We successfully isolated OMVs using standardized protocols and determined their characteristics and concentrations through DLS and BCA protein assay.
- We further engineered the E.coli BL21(DE3) omp8 strain, in order to express the eCPX-SpyTag fusion protein as part of our modular approach for OMV functionalization.
- We proved the presence of plasmids within isolated OMV suspensions.
- We were able to purify recombinant fusion proteins consisting of a phage-tail protein and SpyCatcher, and functionalise them on OMV surfaces, as part of our modular approach for precision-delivery for AMP encoding plasmids.
- We tested AMPs, OMVs and lipid-based nanocarriers for cytotoxicity using the MTT and SYTOX Green assays on lung epithelial cells for safety measures.