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The Riboregulator

The Riboregulator. Method of translational control of gene expression cis -repressive sequence (“lock”) upstream of a gene’s 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.

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The Riboregulator

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  1. The Riboregulator • Method of translational control of gene expression • cis-repressive sequence (“lock”) upstream of a gene’s 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 key3 + 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 we’d 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 it’d be cool if we had multiple pairs built by then so we could show a table or some sort of graphic that shows orthogonality.

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