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

Kyla Bourque, Dominic Devost, Asuka Inoue, Terence E. Hébert

研究成果: Chapter

1 被引用数 (Scopus)

抄録

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.

本文言語English
ホスト出版物のタイトルMethods in Molecular Biology
出版社Humana Press Inc.
ページ169-182
ページ数14
DOI
出版ステータスPublished - 2019

出版物シリーズ

名前Methods in Molecular Biology
1947
ISSN(印刷版)1064-3745
ISSN(電子版)1940-6029

ASJC Scopus subject areas

  • 分子生物学
  • 遺伝学

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