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Team:Alberta NINT/Project
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Manipulating RNA attenuator sequences to generate genetic logic circuits in E. coli...
While electronic devices are inherently connectible due to their common currency (flow of electrons), biological circuits are more complex because they lack connectivity. The Logi-coli[i] project hopes to construct connectible biological circuits based on antisense RNA regulation of transcriptional attenuation. Transcriptional attenuation is a common prokaryotic mechanism for controlling transcriptional productivity. As RNA transcripts are produced by the enzyme RNA polymerase moving along the DNA template strand, they often form secondary stem-loop structures. The terminator is a simple stem-loop followed by the “UUUAUUU” terminator sequence that causes the severing of the nascent RNA strand and the dislocation of RNA polymerase from the DNA template strand. An attenuator is a terminator structure found at the start of a transcript that causes the transcript to be prematurely truncated. Transcriptional attenuation can be modified by designing an anti-sense input RNA strand to utilize base-pairing with complementary parts of the stem-loop structure of the attenuator to disrupt it. In this way, RNA polymerase would pass over the attenuator sequence and continue with transcription. Using software called RNAstructure, our team will be able to rationally design the appropriate input RNA and attenuator sequences to accomplish this. These specific sequences will interact with specified free energies and base-pair complementarity. In this way, the output RNA strand of one circuit can be used as the input RNA strand to the next circuit. This provides a means for connecting a series of circuits. For the final output, a coding region of LacZ or GFP would be transcribed to provide evidence of the connectivity of the circuits. Complex circuits such as AND, OR, NOT, EXOR, and NAND gates can be connected together to create half-adders or full-adders.
Project Details
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