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

Eric M Jones1, Rishi Jajoo1, Daniel Cancilla1

  • 1Department of Chemistry and Biochemistry, UCLA-, DOE Institute for Genomics and Proteomics, Molecular Biology Institute, Quantitative and Computational Biology Institute, Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, and Jonsson Comprehensive Cancer Center, UCLA, Los Angeles, CA 90095, USA.

Cell Systems
|March 25, 2019
PubMed
Summary
This summary is machine-generated.

This study developed a new platform to screen many chemicals against olfactory receptors (ORs), which are G protein-coupled receptors (GPCRs). The platform identified 79 new interactions, including ligands for 15 previously unknown receptors.

Keywords:
GPCRMPRANGShigh-throughputmultiplexolfactionsynthetic biology

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genetics

Background:

  • G protein-coupled receptors (GPCRs) are crucial for cellular chemical sensing.
  • Mammalian olfactory receptors (ORs), the largest GPCR family, are responsible for smell but have many unidentified ligands.
  • Understanding OR-ligand interactions is key to olfactory research.

Purpose of the Study:

  • To develop a high-throughput screening platform for mapping olfactory receptors to odorants.
  • To identify novel ligand-receptor interactions for mammalian olfactory receptors.
  • To characterize previously orphaned olfactory receptors.

Main Methods:

  • Utilized a multiplexed assay with stably engineered human cell lines containing barcoded genetic reporters.
  • Employed next-generation sequencing to analyze reporter activity.
  • Screened 39 mammalian olfactory receptors against a panel of 181 odorants.

Main Results:

  • Identified 79 novel olfactory receptor-odorant interactions.
  • Discovered ligands for 15 olfactory receptors that were previously uncharacterized.
  • Demonstrated the platform's cost-effectiveness and scalability for large-scale chemical library screening.

Conclusions:

  • The developed platform enables efficient and large-scale mapping of chemical libraries to olfactory receptor repertoires.
  • This work significantly expands the known repertoire of olfactory receptor-ligand interactions.
  • Provides a valuable tool for future olfactory research and drug discovery.