Difference between revisions of "Team:William and Mary/Demonstrate"

The central goal of our project this year was to develop a better way for teams to control temporal dynamics by creating an easy to use modular gene expression speed control system. Using a protein degradation tag (pdt) based degradation system, we successfully showed that we can change the speed (time to steady state) for our characterization circuits. While the relationship between degradation and time to steady state had previously been mathematically formulated, we believe that our project represents the first reported biological confirmation of this relationship.

We then expanded on our system by successfully collaborating with University of Maryland iGEM to create a proof of concept version of their copper sensor with increased gene expression speed, and by using our system to create a dynamic circuit, the incoherent feed foreward loop. We believe that we have fulfilled this medal requirement because we have successfully demonstrated that our gene expression speed control system can be used; to modify the time to steady state of an arbitrary protein, as a practical tool, and to create dynamic circuits. Please see our other pages for more background and results. Additionally, see our for judges page and medal requirementsfor an easy way to find all of our pages, and for information on how we fufilled our medal requirements.

Using a series of reporter constructs (see parts for more details), we successfully demonstrated that we could change the speed of gene expression(Figure 1), and that the change was in line with the underlying math model behind our system (Figure 2). We then expanded on this result and showed that as predicted by math modeling, we could successfully alter the gene expression speed of a given circuit while maintaining the steady state value (Figure 3)

Using our pdt system, we were successfully able to show that we could in fact change the gene expression speed of University of Marylands copper sensing circuit (Figure 4). Since University of Marylands circuit uses different proteins, reporter, and different (and more toxic) inducers, we view these initial results as evidence that our system can be used to change the gene expression speed of arbitrary circuits. For more see our collaboration page.

Guided by math modeling, we determined that it would be possible to make an incoherant feed foreward loop, using our existing circuit architecture. We determined that by using inducing our Lon and reporter simultaneously, we could generate pulse behavior from our circuit, further we demonstrated that this effect is dependent on the degradation rate, and that it does not occur in weaker tags or when the concentration of mf-Lon has already reached steady state (Figure 5).