The aim of our project is to build a synthetic organelle based on phase separation as a multifunctional platform. Based on the principle of multivalence and interaction, we fused interactional modules into homo-oligomeric tags (HOtags) to form granules in S. cerevisiae.
We have built spontaneous and induced synthetic organelles by specific interaction modules, so that we can control the formation process by different ways for demands in biological engineering. Then we characterized the kinetics and properties of synthetic organelles theoretically and experimentally. These results confirm the potential of synthetic organelles in synthetic biology.
It inspired us to propose some specific applications of our synthetic organelles, including organization hub, sensor, and metabolism regulator. We have verified the feasibility of them by loading GFP-nanobody module, NAD+ sensor module and carotene production module to the whole system.
We believe that our work has reached the medal requirements of demonstration as we have confirmed that our synthetic organelles can be formed in vivo and deliver a range of functions both for engineering and research due to their amazing properties. The concrete demonstration of the whole platform is shown below. You can see more details of experiments and modeling in our Data Page and Modeling
We visited experts from the College of Chemistry and Molecular Engineering and School of Physics of Peking University, respectively, to learn about the current situation surrounding uranium pollution in the real world and how people could control the situation. After finishing the main work, we presented them with the achievements of the project and got their feedback. (Learn more)
We should reproduce all of the experiments that we have done this summer to make sure the results are credible.
We will optimize the whole strategy to enhance the adsorption efficiency by changing pH, temperature, reaction time of crosslinking and clearance. (The efficiency is only about 60% without further optimization)
According to the results for the adsorption of 13nM uranyl, the polymer network exhibited a good ability in a simulated seawater environment. We could thus also look into other usage scenarios of Uranium Reaper, such as bio-mining and uranium enrichment.
Exchange of the SUP module for other functional proteins. For example, we could integrate proteins which could bind other heavy metals such as mercury so that the polymer network could be used to treat other kinds of pollution as well.
We could assemble enzyme systems behind the SpyTag backbone to create a production plant in vitro. In the protein polymeric network, the concentration of enzymes could be increased and the efficiency of biocatalysis may consequently also be enhanced.
If we optimize the number of SpyTag or SpyCatcher modules per protein monomer, as well as the working concentrations of proteins, we may make protein-3D printing using the Spy Crosslinking Network come true.