Performing Chromophore-Assisted Laser Inactivation in Drosophila Embryos Using GFP
| Autor: | Bruno Monier, Bénédicte Sanson, Anne Pélissier-Monier |
|---|---|
| Přispěvatelé: | Laboratoire de Biologie Cellulaire et Moléculaire du Contrôle de la Prolifération (LBCMCP), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre de Biologie Intégrative (CBI), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
MESH: Gene Expression Laser scanning MESH: Myosin Type II MESH: Drosophila Proteins Confocal [SDV]Life Sciences [q-bio] MESH: Photosensitizing Agents Optogenetics MESH: Chromophore-Assisted Light Inactivation Green fluorescent protein law.invention MESH: Drosophila melanogaster MESH: Animals Genetically Modified 03 medical and health sciences Conditional protein inactivation MESH: Green Fluorescent Proteins Live cell imaging law Microscopy MESH: Microscopy Confocal MESH: Animals MESH: Time-Lapse Imaging Live imaging Chemistry MESH: Genes Reporter Rational design MESH: Embryo Nonmammalian Laser Green fluorescent protein (GFP) 030104 developmental biology Genetic photosensitizer Biophysics Drosophila Chromophore-assisted laser inactivation (CALI) MESH: Fluorescence Recovery After Photobleaching |
| Zdroj: | Methods Mol Biol Methods Mol Biol, 1478, pp.161-176, 2016, ⟨10.1007/978-1-4939-6371-3_8⟩ Methods in Molecular Biology ISBN: 9781493963690 |
| DOI: | 10.1007/978-1-4939-6371-3_8⟩ |
| Popis: | International audience; 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. |
| Databáze: | OpenAIRE |
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