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Characterization of Genetically Encoded FRET Biosensors for Rho-Family GTPases

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Rho GTPases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1821))

Abstract

Genetically encoded FRET-based biosensors are increasingly popular and useful tools for examining signaling pathways with high spatial and temporal resolution in living cells. Here, we show basic techniques used to characterize and to validate single-chain, genetically encoded Förster resonance energy transfer (FRET) biosensors of the Rho GTPase-family proteins. Methods described here are generally applicable to other genetically encoded FRET-based biosensors by modifying the tested conditions to include additional/different regulators and inhibitors, as appropriate for the specific protein of interest.

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Acknowledgments

This work was supported by American Cancer Society Lee National Denim Day Postdoctoral Fellowship PF-15-135-01-CSM (S.D.), Irma T. Hirschl Career Scientist Award (L.H.), and NIH grants: T32GM007491 (V.M.); R01 GM071828 and P01 CA100324 (D.C.); and CA205262 (L.H.). A.M.G. was supported by the Summer Undergraduate Research Program (SURP) of the Albert Einstein College of Medicine, Graduate Division of Biomedical Sciences. Sara K. Donnelly and Veronika Miskolci contributed equally to this work.

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Author notes

  1. Sara K. Donnelly and Veronika Miskolci contributed equally to this work.

Authors and Affiliations

  1. Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY, USA

    Sara K. Donnelly, Alice M. Garrastegui, Dianne Cox & Louis Hodgson

  2. Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY, USA

    Sara K. Donnelly, Dianne Cox & Louis Hodgson

  3. Department of Medical Microbiology and Immunology, University of Wisconsin—Madison, Madison, WI, USA

    Veronika Miskolci

Authors
  1. Sara K. Donnelly
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  2. Veronika Miskolci
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  3. Alice M. Garrastegui
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  4. Dianne Cox
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  5. Louis Hodgson
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Corresponding authors

Correspondence to Dianne Cox or Louis Hodgson .

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Editors and Affiliations

  1. The Hull York Medical School, University of Hull, Hull, United Kingdom

    Francisco Rivero

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Donnelly, S.K., Miskolci, V., Garrastegui, A.M., Cox, D., Hodgson, L. (2018). Characterization of Genetically Encoded FRET Biosensors for Rho-Family GTPases. In: Rivero, F. (eds) Rho GTPases. Methods in Molecular Biology, vol 1821. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8612-5_7

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  • DOI: https://doi.org/10.1007/978-1-4939-8612-5_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8611-8

  • Online ISBN: 978-1-4939-8612-5

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