ArticleBiophysical journal2023
Rho MultiBinder, a fluorescent biosensor that reports the activity of multiple GTPases.
Article in Biophysical journal, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed, 6 citations in OpenAlex.
- RhoG, Rac1 and Cdc42 cooperation in cell protrusion revealed by multiplexed optogenetics and biosensor imaging.bioRxiv : the preprint server for biology · 2026Article
- Allosteric Biosensors Unravel GTPase-Effector Feedback.bioRxiv : the preprint server for biology · 2026Article
- Engineered interfaces in Rac1 and Cdc42 biosensors enhance sensitivity and reduce cell perturbation.Molecular biology of the cell · 2026Article
- Unravelling molecular dynamics in living cells: Fluorescent protein biosensors for cell biology.Journal of microscopy · 2025Review
- Cell migration signaling through the EGFR-VAV2-Rac1 pathway is sustained in endosomes.Journal of cell science · 2025Article
- Excitable Rho dynamics control cell shape and motility by sequentially activating ERM proteins and actomyosin contractility.Science advances · 2024Article
- Celebrating the creative scientific life of Ken Jacobson.Biophysical journal · 2023Article
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Authors and funding
8 authors at 2 institutions in 1 country.
Funding
Abstract
Imaging two or more fluorescent biosensors in the same living cell can reveal the spatiotemporal coordination of protein activities. However, using multiple Förster resonance energy transfer (FRET) biosensors together is challenging due to toxicity and the need for orthogonal fluorophores. Here we generate a biosensor component that binds selectively to the activated conformation of three different proteins. This enabled multiplexed FRET with fewer fluorophores, and reduced toxicity. We generated this MultiBinder (MB) reagent for the GTPases RhoA, Rac1, and Cdc42 by combining portions of the downstream effector proteins Pak1 and Rhotekin. Using FRET between mCherry on the MB and YPet or mAmetrine on two target proteins, the activities of any pair of GTPases could be distinguished. The MB was used to image Rac1 and RhoA together with a third, dye-based biosensor for Cdc42. Quantifying effects of biosensor combinations on the frequency, duration, and velocity of cell protrusions and retractions demonstrated reduced toxicity. Multiplexed imaging revealed signaling hierarchies between the three proteins at the cell edge where they regulate motility.
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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.