Periodic Reporting for period 3 - PArtCell (Physiologically Crowded Artificial Cells for Relevant Drug Screens)
Période du rapport: 2022-07-01 au 2023-12-31
Although crowding is essential, it remains a question of how the biochemical equilibrium in cells is controlled. We aim to map and understand these crowding effects with our project PArtCell (Physiologically Crowded Artificial Cells for Relevant Drug Screens). In addition, our goal is to develop physiologically relevant platforms that can be applied to screen new drugs for treating diseases such as Huntington's in these dense environments. We are developing approaches in natural and artificial cell systems that we will compare to each other to make the most relevant artificial cells possible. Compared to living cells, analyses of their synthetic counterparts have several advantages – for example, natural cells are influenced by many unknown parameters. In artificial cells, on the other hand, the initial conditions can be primarily defined and thus controlled.
We achieved the following milestones:
- We developed a method to better follow the toxic proteins in great detail and in high throughput, which is needed for artificial cell experiments. This method can be applied to multiple (non)toxic proteins and shows the structure of how they stick together.
- We aimed to better map crowding effects in living cells and found that the organization of the molecules should play a major role in the crowding
- We developed a new platform to make artificial cells by microfluidics. This is the first method to do so by microfluidics, where crowded minimal-oil-containing vesicles are obtained.
We show for the first time that cell wall damage can increase crowding in bacterial cells, providing the clearest indication thus far that cytoplasmic organization influences crowding.
In our artificial cell production, we developed the first microfluidic method to make these with high internal crowding and minimal additional oil present.
We expect to further these artificial cells as drug screening platforms, in which we have proteins in their toxic form induced by their crowded environment.