ArticleScientific reports2017
Using nanoBRET and CRISPR/Cas9 to monitor proximity to a genome-edited protein in real-time.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed.
- A calcium-sensing receptor dileucine motif directs internalization to spatially distinct endosomal signaling pathways.iScience · 2025Article
- Bioluminescence Resonance Energy Transfer (BRET) Assay to Measure Gq Recruitment to the Ghrelin Receptor.Methods in molecular biology (Clifton, N.J.) · 2025Article
- CXCL17 is an allosteric inhibitor of CXCR4 through a mechanism of action involving glycosaminoglycans.Science signaling · 2024Article
- Article
- Elucidating the Interactome of G Protein-Coupled Receptors and Receptor Activity-Modifying Proteins.Pharmacological reviews · 2023Review
- Detecting and measuring of GPCR signaling - comparison of human induced pluripotent stem cells and immortal cell lines.Frontiers in endocrinology · 2023Review
- The therapeutic potential of GLP-1 receptor biased agonism.British journal of pharmacology · 2022 · on this mapReview
- GPCR heteromers: An overview of their classification, function and physiological relevance.Frontiers in endocrinology · 2022Review
- CRISPR/Cas9 Genome Editing vs. Over-Expression for Fluorescent Extracellular Vesicle-Labeling: A Quantitative Analysis.International journal of molecular sciences · 2021Article
- Profiling novel pharmacology of receptor complexes using Receptor-HIT.Biochemical Society transactions · 2021Review
- Detection of genome-edited and endogenously expressed G protein-coupled receptors.The FEBS journal · 2021Review
- Investigation of Receptor Heteromers Using NanoBRET Ligand Binding.International journal of molecular sciences · 2021Article
- A nanoluciferase biosensor to investigate endogenous chemokine secretion and receptor binding.iScience · 2021Article
- NanoLuc-Based Methods to Measure β-Arrestin2 Recruitment to G Protein-Coupled Receptors.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Ligand-directed covalent labelling of a GPCR with a fluorescent tag in live cells.Communications biology · 2020Article
- CRISPR-Mediated Protein Tagging with Nanoluciferase to Investigate Native Chemokine Receptor Function and Conformational Changes.Cell chemical biology · 2020Article
- Importance of Quantifying Drug-Target Engagement in Cells.ACS medicinal chemistry letters · 2020Article
- Fluorescent ligands: Bringing light to emerging GPCR paradigms.British journal of pharmacology · 2020Review
- Optimised insert design for improved single-molecule imaging and quantification through CRISPR-Cas9 mediated knock-in.Scientific reports · 2019Article
- Complex Formation between VEGFR2 and the βCell chemical biology · 2019Article
Corrections and comments
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Bioluminescence resonance energy transfer (BRET) has been a vital tool for understanding G protein-coupled receptor (GPCR) function. It has been used to investigate GPCR-protein and/or -ligand interactions as well as GPCR oligomerisation. However the utility of BRET is limited by the requirement that the fusion proteins, and in particular the donor, need to be exogenously expressed. To address this, we have used CRISPR/Cas9-mediated homology-directed repair to generate protein-Nanoluciferase (Nluc) fusions under endogenous promotion, thus allowing investigation of proximity between the genome-edited protein and an exogenously expressed protein by BRET. Here we report BRET monitoring of GPCR-mediated β-arrestin2 recruitment and internalisation where the donor luciferase was under endogenous promotion, in live cells and in real time. We have investigated the utility of CRISPR/Cas9 genome editing to create genome-edited fusion proteins that can be used as BRET donors and propose that this strategy can be used to overcome the need for exogenous donor expression.
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