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Immune Mediators in Cancer pp 281–293Cite as

Optogenetics: Rho GTPases Activated by Light in Living Macrophages

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2108))

Abstract

Genetically encoded optogenetic tools are increasingly popular and useful for perturbing signaling pathways with high spatial and temporal resolution in living cells. Here, we show basic procedures employed to implement optogenetics of Rho GTPases in a macrophage cell line. Methods described here are generally applicable to other genetically encoded optogenetic tools utilizing the blue-green spectrum of light for activation, designed for specific proteins and enzymatic targets important for immune cell functions.

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Acknowledgements

This work was supported by National Institutes of Health grant T32GM007491 to V.M., R01GM129098 and R01GM132098 to L.H. National Cancer Institute P30CA013330 for Analytical Imaging Facility support. Irma T. Hirschl Career Scientist Award to L.H. pTriEX-mVenus-PA-Cdc42 (Addgene #75263), pTriEX-mVenus-PA-Rac1 (Addgene #22007), pTriEX-PA-Rac1 C450R (Addgene #22025), pTriEX-NTOM20-LOV2 WT (Addgene #81009) and pTriEx-mVenus-Zdk1-VAV2 DH/PH/C1 (Addgene #81133) were gifts from Dr. Klaus Hahn (UNC-Chapel Hill).

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Authors and Affiliations

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

    Maren Hülsemann, Polina V. Verkhusha, Peng Guo, Dianne Cox & Louis Hodgson

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

    Maren Hülsemann, Peng Guo, Dianne Cox & Louis Hodgson

  3. Analytical Imaging Facility, Albert Einstein College of Medicine, Bronx, NY, USA

    Peng Guo

  4. Department of Medical Microbiology and Immunology, University of Wisconsin–Madison, Madison, WI, USA

    Veronika Miskolci

Authors
  1. Maren Hülsemann
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  2. Polina V. Verkhusha
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  3. Peng Guo
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  4. Veronika Miskolci
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  5. Dianne Cox
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  6. Louis Hodgson
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Corresponding authors

Correspondence to Dianne Cox or Louis Hodgson .

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Editors and Affiliations

  1. Department of Biological Sciences, St. John’s University, Queens, NY, USA

    Ivana Vancurova

  2. Department of Biological Sciences, St. John’s University, Queens, NY, USA

    Yan Zhu

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Hülsemann, M., Verkhusha, P.V., Guo, P., Miskolci, V., Cox, D., Hodgson, L. (2020). Optogenetics: Rho GTPases Activated by Light in Living Macrophages. In: Vancurova, I., Zhu, Y. (eds) Immune Mediators in Cancer. Methods in Molecular Biology, vol 2108. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0247-8_24

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  • DOI: https://doi.org/10.1007/978-1-0716-0247-8_24

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0246-1

  • Online ISBN: 978-1-0716-0247-8

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