1. A Scalable, Multiplexed Assay for Decoding GPCR-Ligand Interactions with RNA Sequencing
- Author
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Daniel Cancilla, Sriram Kosuri, Jeffrey C. Wang, Joshua S. Bloom, Hiroaki Matsunami, Nathan B. Lubock, Rishi Jajoo, Rocky Cheung, Eric M. Jones, Claire A. de March, and Megan Satyadi
- Subjects
Histology ,Computational biology ,Olfaction ,Biology ,Ligands ,Receptors, Odorant ,Article ,Pathology and Forensic Medicine ,Cell Line ,03 medical and health sciences ,Synthetic biology ,0302 clinical medicine ,Animals ,Humans ,Multiplex ,Receptor ,030304 developmental biology ,G protein-coupled receptor ,Mammals ,0303 health sciences ,Ligand ,Sequence Analysis, RNA ,Gene Expression Profiling ,RNA ,Cell Biology ,Small molecule ,Smell ,Odorants ,030217 neurology & neurosurgery ,Signal Transduction - 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 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 olfactory receptors 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.
- Published
- 2018