IGEM:Stanford/2010/Notebook/mRNA sRNA
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General Ideas
One Idea to measure [B] / [A] (assume [A] ≠ 0)
- input A induces expression of GFP mRNA
- input B induces expression of GFP sRNA (sRNA that targets the GFP mRNA transcript)
- so for a given [A],
- if [B] = 0, we should see glowing bacteria
- if 0 < [B] < [A], we should see some glow
- if [B] > [A], we should see no glow
Another Idea to measure [B] / [A] (assume [A] ≠ 0)
- input A induces expression of GFP mRNA and YFP sRNA
- input B induces expression of YFP mRNA and GFP sRNA
- so for a given [A],
- if [B] = 0, we should see GREEN glowing bacteria
- if 0 < [B] < [A], we should see some glow that is more GREEN than YELLOW.
- if [B] > [A], we should see some glow that is more YELLOW than GREEN.
References
- here's a good reference: Engineering RNA-based circuits. It discusses different methods of regulation by RNA:
- via base-pairing (small RNA)
- shape-based (riboswitches)
- catalytic (ribozymes)
- engineered riboregulators (possible mechanism for us) Engineered riboregulators enable post-transcriptional control of gene expression
- Note: apparently does not work well in practice
- engineering those aforementioned sRNA's:
- vague article Engineers meet small RNA
- some specifics Quantitative Characteristics of Gene Regulation by Small RNA