Combining Conformational Profiling of GPCRs with CRISPR/Cas9 Gene Editing Approaches

  • Kyla Bourque
  • Dominic Devost
  • Asuka Inoue
  • Terence E. HébertEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1947)


Ligand-biased signaling could have a significant impact on drug discovery programs. As such, many approaches to screening now target a larger section of the signaling responses downstream of an individual G protein-coupled receptor (GPCR). Biosensor-based platforms have been developed to capture signaling signatures. Despite the ability to use such signaling signatures, they may still be particular to an individual cell type and thus such platforms may not be portable from cell to cell, necessitating further cell-specific biosensor development. We have developed a complementary strategy based on capturing receptor-proximal conformational profiles using intra-molecular BRET-based sensors composed of a Renilla luciferase donor engineered into the carboxy-terminus and CCPGCC motifs which bind fluorescent hairpin biarsenical dyes engineered into different positions into the receptor primary structure. Here, we discuss how these experiments can be conducted and combined with CRISPR/Cas9 genome editing to assess specific G protein-dependent and -independent events.

Key words

GPCRs BRET Genome-editing Conformation-sensitive biosensors CRISPR Drug discovery 



The work was funded by a grant to TEH from the Canadian Institutes for Health Research (MOP-130309). KB was supported by an internal Faculty of Medicine Studentship from McGill University.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Kyla Bourque
    • 1
  • Dominic Devost
    • 1
  • Asuka Inoue
    • 2
    • 3
  • Terence E. Hébert
    • 1
    Email author
  1. 1.Department of Pharmacology and TherapeuticsMcGill UniversityMontrealCanada
  2. 2.Graduate School of Pharmaceutical SciencesTohoku UniversitySendaiJapan
  3. 3.Japan Science and Technology Agency (JST)Precursory Research for Embryonic Science and Technology (PRESTO)KawaguchiJapan

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