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Drosophila pp 161–176Cite as

Performing Chromophore-Assisted Laser Inactivation in Drosophila Embryos Using GFP

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1478))

Abstract

Chromophore-assisted laser inactivation (CALI) is an optogenetic technique in which light-induced release of reactive oxygen species triggers acute inactivation of a protein of interest, with high spatial and temporal resolution. At its simplest, selective protein inactivation can be achieved via the genetic fusion of the protein to a photosensitizer such as EGFP, and using standard optical setups such as laser scanning confocal microscopes. Although use of CALI in Drosophila is relatively recent, this technique can be a powerful complement to developmental genetics, especially in vivo as it allows visualization of the immediate consequences of local protein inactivation when coupled to time-lapse microscopy analysis. In addition to providing examples of protocols, this chapter is intended as a conceptual framework to support the rational design of CALI experiments.

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Acknowledgements

We thank Andrew Davidson and Will Wood for critical reading of the manuscript. BS acknowledges funding from a Welcome Trust Investigator Award [099234/Z/12/Z]. APM and BM are supported respectively by Université Toulouse III-Paul Sabatier and Centre National de la Recherche Scientifique (CNRS).

Author information

Authors and Affiliations

  1. Université Toulouse III-Paul Sabatier, 118 route de Narbonne, 31062, Toulouse, France

    Anne Pélissier-Monier & Bruno Monier

  2. CNRS, UMR5547, Centre de Biologie du Développement, 31062, Toulouse, France

    Anne Pélissier-Monier

  3. Department of Physiology, Development and Neuroscience, University of Cambridge, Anatomy Building, Downing Street, Cambridge, CB2 3DY, UK

    Bénédicte Sanson

  4. Laboratoire de Biologie Cellulaire et Moléculaire du Contrôle de la Prolifération, CNRS, UMR5088, 31062, Toulouse, France

    Bruno Monier

Authors
  1. Anne Pélissier-Monier
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  2. Bénédicte Sanson
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  3. Bruno Monier
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Corresponding author

Correspondence to Bénédicte Sanson .

Editor information

Editors and Affiliations

  1. Institute of Genetics, Technische Universität Dresden, Dresden, Germany

    Christian Dahmann

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Pélissier-Monier, A., Sanson, B., Monier, B. (2016). Performing Chromophore-Assisted Laser Inactivation in Drosophila Embryos Using GFP. In: Dahmann, C. (eds) Drosophila. Methods in Molecular Biology, vol 1478. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6371-3_8

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  • DOI: https://doi.org/10.1007/978-1-4939-6371-3_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6369-0

  • Online ISBN: 978-1-4939-6371-3

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