Fig. 1. Design of protease-based live yeast biosensor. (IMAGE)
Caption
(A) Engineered yeast has a GPCR that induces readout expression in response to cognate peptide. In addition, a peptide protease is constitutively secreted by engineered yeast to cleave the cognate peptide. (B) Peptide dose–response plot illustrating the expected effect of the secreted protease. Peptide cleavage by the protease reduces GPCR activation as observed by the apparent shift in EC50. (C) Dose–response curve characterizing the readout for distinction of 2 peptide variants within expected peptide concentration range (gray box). Without protease, the GPCR response to both peptides should show similar activation curve. With protease, only one peptide is cleaved and will have shifted apparent EC50 outside gray box. (D) Illustration of a paper-based dipstick assay with engineered yeast patches responding to peptides differing in single-amino-acid (columns). The rows contain yeast expressing (+protease) or not expressing (−protease) the peptide protease. The orange pigment represents an inducible readout that can distinguish peptide variants. A.U., arbitrary units.
Credit
BioDesign Research
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