IGEM:Stanford/2010/Notebook/mRNA sRNA

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