Experiment:
1. We constructed a part library that can drive protein phase separation. The library includes several dimerization modules and multivalent modules.
2. We rationally designed a synthetic organelle called SPOT (Synthetic Phase separation-based Organelle Platform) and used it to achieve multi-functions. We believed SPOT to be a useful platform for other iGEM teams to investigate protein phase separation and design synthetic organelles.
3. The synthetic organelles could form spontaneously in yeast. They could also be induced by rapamycin. We chose different platforms according to the different functions we wanted to achieve.
4. We set up a thermodynamic model and a kinetic model to characterize our system. The experimental results were consistent with the models.
5. We tested the strength of four different yeast promoters using flow cytometry.
6. We thoroughly tested some key parameters of the paired FKBP-HOTag3 and Frb-HOTag6. The thermodynamic properties and the kinetic properties were well characterized.
7. We explored the biophysical properties of the SUMO-SIM system. We proved the granules were liquid-like.
8. Three enzymes were essential for β-carotene producing in yeast. We loaded the three enzymes on the synthetic organelle and increased the reaction rate . We confirmed SPOT could function as a reaction crucible.
9. The inducible SPOT could sequester rapamycin with high efficiency. We used SPOT as a cure of the toxicity of rapamycin.
10. We verified the high sensibility of SPOT-based sensor system. We also built an ABA sensor.
Beyond experiment:
1. We submitted 29 high-quality Biobricks, including basic parts, such as multivalent HOTags, and composite parts that can robustly drive phase separation.
2. We developed a batch file to process large amounts of fluorescence images. It offered a high throughput solution to analyze experimental results derived from fluorescence microscope.
3. We did a statistical investigation of iGEM in the past decade. The academic background of the iGEMers provided evidence of the increasing diversity of iGEM.
4. We played an active part in preschool education and tried to integrate some biological knowledge into the games. We also devoted ourselves to promoting synthetic biology in high schools.
5. We focused on how to build a bridge between scientific research and the public. We exchanged ideas with experts from academia and developed a feasible idea to design a cheap fluorescence microscope affordable by the public.
6. We collaborated with many universities. We shared experimental equipment and helped each other with modeling as well as experiments. We also attended CCiC (Conference of China iGEMer Community) and got inspiration from other teams.