Which Output Though?!

The wnt pathway is finally coming together (somewhat).  We are beginning to debug our original system in the hopes of finding a more efficient and reliable system.  Finding a way to reduce/eliminate false negatives, exploring proper marking, signaling, and output, as well as simply improving the pathway are our priorities.

I have been focused on the output stage of the wnt pathway, trying to figure out the most plausible output mechanism.  Originally, we went with generic GFP as it seemed to do everything what we wanted, and most importantly was easy to analyze.  The introduction of additional genomic sequences or markers then altered this original idea.

We chose to include topflash in our design, which uses luciferase as an output instead of GFP.  Luciferase has many benefits over GFP, including most importantly, the better ability to be amplified.  One of the problems we have is the fact that if only a few cells are detected, the fluorescent output may be hard to analyze.  Amplifying the output will solve this problem, and using an output that can be easily manipulated such as luciferase is to our benefit.

There is still a lot to do, but what we have accomplished so far is quite astonishing.  I can't wait to see what will be the eventual outcome of our project, and what the next steps will be.