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Visualizing endogenous Rho activity with an improved localization-based, genetically encoded biosensor.

Eike K Mahlandt1, Janine J G Arts1,2, Werner J van der Meer1

  • 1Swammerdam Institute for Life Sciences, Section of Molecular Cytology, van Leeuwenhoek Centre for Advanced Microscopy, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands.

Journal of Cell Science
|August 6, 2021
PubMed
Summary
This summary is machine-generated.

Researchers developed improved Rho biosensors for real-time monitoring of Rho GTPase activity. These genetically encoded fluorescent probes enhance understanding of cellular processes like migration and division.

Keywords:
BiosensorEndothelial cellRhoRho GTPaseThrombin

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Rho GTPases regulate crucial cell functions including morphology, polarity, and migration.
  • Understanding Rho GTPase activity is vital for studying development and cell migration.
  • Localization-based Rho biosensors offer real-time, subcellular insights into Rho GTPase activity in living systems.

Purpose of the Study:

  • To systematically evaluate and improve localization-based Rho biosensors for detecting endogenous Rho activity.
  • To characterize the impact of fluorescent proteins and Rho-binding peptides on biosensor performance.
  • To enable broader applications of Rho relocation biosensors in various cellular contexts.

Main Methods:

  • Systematic examination of fluorescent protein and Rho-binding peptide contributions to sensor performance.
  • Comparison of relocation efficiency and specificity using cell-based assays.
  • Development and validation of genetically encoded fluorescent biosensors.

Main Results:

  • Identification of several improved localization-based, genetically encoded fluorescent biosensors for Rho activity.
  • Demonstrated broader applicability of the new biosensors in visualizing Rho activity during cell division, migration, and GPCR signaling.
  • Enhanced avidity of new biosensors for Rho activity.

Conclusions:

  • The developed Rho relocation biosensors offer improved performance and broader applicability for studying Rho GTPase-mediated cellular processes.
  • These enhanced biosensors facilitate a deeper understanding of cellular functions regulated by Rho.
  • The study paves the way for more effective real-time monitoring of Rho activity in diverse biological contexts.