Future works

Cyclebow experiments

Although we have successfully constructed our Cyclebow, it is only a prototype that tracks the limited number of cell-cycles. In addition, one year is far from enough for a convincing and rigorous project, there are many aspects and details during the experiments that we intended to analyze, yet didn’t have the time. For instance, we didn’t confirm the half-life of the fluorescences, which might be a concern for the color distinguishment.

Moreover, in our design, the COUNTER CIRCUIT should consist VIKA UNIT and VCRE UNIT, and our REPORTER UNIT should contain red, green and blue. We suspended this design for the reason that VCRE UNIT and GFP UNIT were not available.

Hybrid-color Cyclebow

In our initial design, we use pure color for distinguishing cell-cycles (e.g., red of mCherry, green of GFP). Having three different pure colors, we can only track three cell-cycles, and those that experienced three or more will all be presenting the same color.

To achieve the tracking for extended cell cycles, we devised a plan of using “hybrid colors”. By duplicating copy number of the Circuit Reporter, we create the possibility of not cutting out all the same florescence UNIT. Therefore, instead of pure colors, cells that undergo different numbers of cell-cycles will present somewhat a continuous spectrum of which color ranges from the first UNIT’s color to the last UNIT’s color.

Figure 1: Concept of the outcome of Hybrid-color cyclebow
Cited from Kretzschmar, K., & Watt, F. M. (2012). Lineage tracing. Cell, 148(1-2), 33-45.
doi: 10.1016/j.cell.2012.01.002

This part is detailedly demonstrated in our Cyclebow Hybrid-color model.

Tests in animal models

Although our prototype was constructed in tumor cells, our intention goes for stem cells such as MSCs. Therefore, Cyclebow could be further induced in stem cells, and the stem cells can be implanted into animal models, such as mice. We are confident that the experimental data will provide insight for stem cell therapy.