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A Scalable, Multiplexed Assay for Decoding GPCR-Ligand Interactions with RNA Sequencing.


ABSTRACT: G protein-coupled receptors (GPCRs) are central to how mammalian cells sense and respond to chemicals. Mammalian olfactory receptors (ORs), the largest family of GPCRs, mediate the sense of smell through activation by small molecules, though for most bonafide ligands, they have not been identified. Here, we introduce a platform to screen large chemical panels against multiplexed GPCR libraries using next-generation sequencing of barcoded genetic reporters in stably engineered human cell lines. We mapped 39 mammalian ORs against 181 odorants and identified 79 interactions that have not been reported to our knowledge, including ligands for 15 previously orphaned receptors. This multiplexed receptor assay allows the cost-effective mapping of large chemical libraries to receptor repertoires at scale.

SUBMITTER: Jones EM 

PROVIDER: S-EPMC6907015 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature

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A Scalable, Multiplexed Assay for Decoding GPCR-Ligand Interactions with RNA Sequencing.

Jones Eric M EM   Jajoo Rishi R   Cancilla Daniel D   Lubock Nathan B NB   Wang Jeffrey J   Satyadi Megan M   Chong Rockie R   de March Claire C   Bloom Joshua S JS   Matsunami Hiroaki H   Kosuri Sriram S  

Cell systems 20190320 3


G protein-coupled receptors (GPCRs) are central to how mammalian cells sense and respond to chemicals. Mammalian olfactory receptors (ORs), the largest family of GPCRs, mediate the sense of smell through activation by small molecules, though for most bonafide ligands, they have not been identified. Here, we introduce a platform to screen large chemical panels against multiplexed GPCR libraries using next-generation sequencing of barcoded genetic reporters in stably engineered human cell lines. W  ...[more]

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