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3) Inhibitor aptamer RNA product successfully suppresses the transcriptional function of their specific target enzyme.
3) Inhibitor aptamer RNA product successfully suppresses the transcriptional function of their specific target enzyme.
 
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Gel electrophoresis is another useful tool in determining the functionality of enzymes prior and post interaction of the inhibiting aptamer RNA products.
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<u>DNA Strands Binding to RNA Aptamers</u>
<u>DNA Strands Binding to RNA Aptamers</u>

Revision as of 23:25, 19 October 2014

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Results



ODE Models


Aptamer-Kleptamer Pairs

The oligonucleotide sequences are specified in the Supplementary section. Once the necessary genelets and strands for our RNA clocks and switches systems are designed, it is important to characterize and verify that our DNA sequences are designed correctly.

Inhibition of SP6 RNA Polymerase.
Inhibition of T7 RNA Polymerase.

Enzyme Inhibition with RNA Aptamers
The topologies of our RNA clocks and switches rely on the idea that inhibition of a module is possible, whether it is self-mediated or caused by another module. A fluorescent RNA aptamer genelets, Malachite Green and Spinach, are used to characterize and quantify our system. This reporting system will be our visual determining guide for whether the enzyme's transcriptional funciton is altered, specifically inhibition. A spectrofluorometer is used to characterize the inhibition of T7 RNA Polymerase and SP6 RNA Polymerase by measuring fluorescence in the system. These experiments successfully demonstrated three important concepts that are necessary to achieve oscillations and bistablity behaviors for our systems:
1) The proper binding of specific enzymes to the inhibiting aptamer sequences.
2) The production of RNA aptamer strands through transcription of DNA to RNA.
3) Inhibitor aptamer RNA product successfully suppresses the transcriptional function of their specific target enzyme.
Gel electrophoresis is another useful tool in determining the functionality of enzymes prior and post interaction of the inhibiting aptamer RNA products.

DNA Strands Binding to RNA Aptamers


Aptamer-Kleptamer Interaction



Assembling the Circuits


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