The Riboregulator

1
The Riboregulator

• Method of translational control of gene
  expression
• cis-repressive sequence (lock) upstream of a
  genes coding region forms a hairpin,
  sequestering the ribosome binding site
• trans-activating (key) mRNA strand binds and
  opens the hairpin thus allowing access to the RBS.

• Highly specific activation occurs. Very similar
  lock and key pair sequences do not exhibit
  crosstalk

Isaacs et al., Nature Biotechnology, 2004
2
Results with lock3/key3
Strain Fluorescence no plasmids 31 lock3RFP 44 key
3 lock3RFP 78 OnRFP 6415
key3

lock3-RFP
3
Improved locks and keys
Frame of homology
Presence of hairpin
Position of promoter
Degree of homology
Position of terminator Transcriptional fusion
Length of spacer
Distance from RBS
4
New key architectures

• Increases effective key concentration

• Increases stability

5
This is where wed have the best lock and key
pair detailed with specific dimensions and good
assay data (e.g., FACS, QPCR, OD/flourescense
time couse, et cetera). And itd be cool if we
had multiple pairs built by then so we could show
a table or some sort of graphic that shows
orthogonality.