BI-STABLE CIRCUIT DESIGN (6 points possible)(Challenging) We are now ready to choose the biological parts to implement an abstract GRN! You will use one of the GRNs that you found suitable in the previous exercise as the basis for this exercise. To figure out the parts you're going to use, you look into your parts library and find the following biological components:
With a subset of these parts, construct a two state circuit (mKate on, GFP off; mKate off, GFP on) by dragging the parts onto the DNA circuit diagram below. The inputs to your circuit should be dox and shFF4. Note:
Your 3rd point is not entirely correct. Question 3 does have some circuits where dox is as you an "activating inducer". Any circuit where the SM is acting on a repressor is what you would call an "activating inducer" (there are 4). Also, a better term for doxycycline (dox) in this case is a derepressorof TetR promoters. It serves to remove repression of TetO by TetR.
That being said, you're 4th point is a subtle, but very important one. Whenever using miRNAs to compute logic, one must think about the mechanism behind it. Due to the spatiotemporal separation of RNA processing and miRNA repression, it turns out miRNA cannot be used to repress another miRNA! |
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