Bioluminescence resonance energy transfer (BRET) is widely employed for real-time monitoring of G protein-coupled receptor activity, interactions, and trafficking in heterologous cell lines, yet its use in neuronal systems remains limited. Here, we present a protocol to apply BRET assays to primary neuronal cultures from mouse embryos. We describe steps and key concepts for generating plasmid constructs and lentivirus preparations, plating and lentiviral transduction of primary cultured neurons in 96-well plates, and BRET data collection and analysis. For complete details on the use and execution of this protocol, please refer to George et al..
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http://dx.doi.org/10.1016/j.xpro.2024.103228 | DOI Listing |
J Enzyme Inhib Med Chem
December 2025
Laboratory of Biochemistry and Molecular Biology, Department of Chemistry, Life Sciences and Environmental Sustainability, University of Parma, Parma, Italy.
Antibiotic resistance stands as the foremost post-pandemic threat to public health. The urgent need for new, effective antibacterial treatments is evident. Protein-protein interactions (PPIs), owing to their pivotal role in microbial physiology, emerge as novel and attractive targets.
View Article and Find Full Text PDFBiosensors (Basel)
November 2024
Department of Pathology and Molecular Medicine, Queen's University, Kingston, ON K7L 3N6, Canada.
Recent advances in drug discovery have established biosensors as indispensable tools, particularly valued for their precision, sensitivity, and real-time monitoring capabilities. The review begins with a brief overview of cancer drug discovery, underscoring the pivotal role of biosensors in advancing cancer research. Various types of biosensors employed in cancer drug discovery are then explored, with particular emphasis on fluorescence- and bioluminescence-based technologies such as FRET, TR-FRET, BRET, NanoBRET, and NanoBiT.
View Article and Find Full Text PDFbioRxiv
December 2024
Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, La Jolla, CA, USA.
The constitutive (ligand-independent) signaling of G protein-coupled receptors (GPCRs) is being increasingly appreciated as an integral aspect of their function; however, it can be technically hard to detect for poorly characterized, e.g. orphan, receptors of the cAMP-inhibitory Gi-coupled (GiPCR) family.
View Article and Find Full Text PDFExtracell Vesicles Circ Nucl Acids
December 2023
School of Bioscience and Technology, Weifang Medical University, Weifang 261053, Shandong, China.
Cells have the capability to discharge extracellular vesicles (EVs) into a range of bodily fluids. Extracellular vesicles (EVs) encapsulate biological molecules such as proteins and nucleic acids, playing a role in facilitating cell-cell communication. They actively engage in a myriad of physiological and pathological processes.
View Article and Find Full Text PDFBr J Pharmacol
December 2024
INSERM UMR1149/Inflammation Research Center (CRI), Team "From Inflammation to Cancer in Digestive diseases (INDiD)", DHU UNITY, Université Paris Cité, Paris, France.
Background And Purpose: Orexins have important biological effects on the central and peripheral nervous systems. Their primary ability is to regulate the sleep-wake cycle. Orexins and their antagonists, via OX receptor have been shown to have proapoptotic and antitumor effects on various digestive cancers cell models.
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