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Heterologous Expression of Olfactory Receptors for Targeted Chemosensing
Author(s) -
Dhanasekaran Danny N.,
Radhika Venkat,
ProikasCezanne Tassula,
Jayaraman Muralidharan,
Ha Jihee
Publication year - 2009
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1111/j.1749-6632.2009.04374.x
Subject(s) - heterologous , receptor , biology , yeast , olfactory system , heterologous expression , olfactory receptor , microbiology and biotechnology , saccharomyces cerevisiae , signal transduction , neuroscience , computational biology , biochemistry , gene , recombinant dna
With the broad objective of developing a heterologous expression system for the mammalian olfactory signaling pathway, we have engineered yeast cells in which the mammalian olfactory signaling pathway is genetically integrated. Our results demonstrate that the prototypic “olfactory yeast” strain WIF‐1α can sense and report the presence of defined chemical agents through the engineered mammalian olfactory system. In this heterologous Saccharomyces cerevisiae ‐based expression system, the primary components of the mammalian olfactory signaling pathway have been engineered, and signaling by the rat olfactory receptor is coupled to the expression of green fluorescent protein. By shuttling a library of olfactory receptor ligand‐binding pockets into the pre‐engineered signaling units of WIF‐1 yeast cells, we further demonstrate the ability of these olfactory yeast cells to detect 2,4‐dinitrotoluene. Using this approach, our results have identified the novel rat olfactory receptor Olfr226 as a 2,4‐dinitrotoluene‐responsive receptor. Genetic integration of a highly discriminatory olfactory system into biologically stable and biochip‐adaptable yeast cells, as presented here, can provide an ideal targeted chemosensing platform for detecting diverse chemical molecules. In addition to their potential use in deorphanizing the superfamily of olfactory receptors, the engineered olfactory yeast cells should be amenable for high‐throughput screening to identify receptor‐specific molecular targets.