ArticleAntibodies (Basel, Switzerland)2019
Selection and Characterization of a Nanobody Biosensor of GTP-Bound RHO Activities.
Article in Antibodies (Basel, Switzerland), 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
What it found
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Who cites it
14 citing papers in PubMed, 28 citations in OpenAlex.
- Intracellular protein binders for imaging, control and future therapeutics.Nature biomedical engineering · 2026Review
- Nanobodies: From Discovery to AI-Driven Design.Biology · 2025Review
- Electrochemical and Photoelectrochemical Immunosensors for the Detection of Ovarian Cancer Biomarkers.Sensors (Basel, Switzerland) · 2023Review
- mNG-tagged fusion proteins and nanobodies to visualize tropomyosins in yeast and mammalian cells.Journal of cell science · 2022Article
- Affimers and nanobodies as molecular probes and their applications in imaging.Journal of cell science · 2022Article
- Seeing is believing: tools to study the role of Rho GTPases during cytokinesis.Small GTPases · 2022Review
- Intracellular VHHs to monitor and modulate GPCR signaling.Frontiers in endocrinology · 2022Review
- Tripartite split-GFP assay to identify selective intracellular nanobody that suppresses GTPase RHOA subfamily downstream signaling.Frontiers in immunology · 2022Article
- Visualizing endogenous Rho activity with an improved localization-based, genetically encoded biosensor.Journal of cell science · 2021Article
- Article
- An Inside Job: Applications of Intracellular Single Domain Antibodies.Biomolecules · 2020Review
- Exploring cellular biochemistry with nanobodies.The Journal of biological chemistry · 2020Review
- Special Issue: Nanobody.Antibodies (Basel, Switzerland) · 2020Article
- Article
Corrections and comments
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Authors and funding
7 authors at 2 institutions in 2 countries.
Funding
Abstract
RHO (Ras HOmologous) GTPases are molecular switches that activate, in their state bound to Guanosine triphosphate (GTP), key signaling pathways, which involve actin cytoskeleton dynamics. Previously, we selected the nanobody RH12, from a synthetic phage display library, which binds the GTP-bound active conformation of RHOA (Ras Homologous family member A). However, when expressed as an intracellular antibody, its blocking effect on RHO signaling led to a loss of actin fibers, which in turn affected cell shape and cell survival. Here, in order to engineer an intracellular biosensor of RHOA-GTP activation, we screened the same phage nanobody library and identified another RHO-GTP selective intracellular nanobody, but with no apparent toxicity. The recombinant RH57 nanobody displays high affinity towards GTP-bound RHOA/B/C subgroup of small GTPases in vitro. Intracellular expression of the RH57 allowed selective co-precipitation with the GTP-bound state of the endogenous RHOA subfamily. When expressed as a fluorescent fusion protein, the chromobody GFP-RH57 was localized to the inner plasma membrane upon stimulation of the activation of endogenous RHO. Finally, the RH57 nanobody was used to establish a BRET-based biosensor (Bioluminescence Resonance Energy Transfer) of RHO activation. The dynamic range of the BRET signal could potentially offer new opportunities to develop cell-based screening of RHOA subfamily activation modulators.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.