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

Cell Syst. 2019 Mar 27;8(3):254-260.e6. doi: 10.1016/j.cels.2019.02.009. Epub 2019 Mar 20.

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.

Keywords: GPCR; MPRA; NGS; high-throughput; multiplex; olfaction; synthetic biology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Line
  • Gene Expression Profiling
  • Humans
  • Ligands
  • Mammals / metabolism
  • Mammals / physiology
  • Odorants*
  • Receptors, Odorant / metabolism*
  • Sequence Analysis, RNA / methods*
  • Signal Transduction*
  • Smell*

Substances

  • Ligands
  • Receptors, Odorant