Team:Stanford/Notebook/mRNA sRNA
From 2010.igem.org
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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: [http://www.springerlink.com/content/x6p4lqn1608267r1/fulltext.pdf 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) [http://www.nature.com/nbt/journal/v22/n7/pdf/nbt986.pdf Engineered riboregulators enable post-transcriptional control of gene expression]
- Note: apparently does not work well in practice
- engineering those aforementioned sRNA's:
- vague article [http://www.nature.com/nmeth/journal/v4/n12/full/nmeth1207-986b.html Engineers meet small RNA]
- some specifics [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1994261/pdf/pbio.0050229.pdf Quantitative Characteristics of Gene Regulation by Small RNA]