Reversible Optogenetic Control of Subcellular Protein Localization in a Live Vertebrate Embryo
| Autor: | Rachel E. Moore, Anna Reade, Orion D. Weiner, Jonathan D.W. Clarke, Anna R. Goldberg, Clare E. Buckley |
|---|---|
| Přispěvatelé: | Buckley, Clare [0000-0003-3329-3973], Apollo - University of Cambridge Repository |
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Technology Light Biology Optogenetics Medical and Health Sciences General Biochemistry Genetics and Molecular Biology 03 medical and health sciences medicine Animals Molecular Biology Zebrafish phytochrome Pard3 asymmetric inheritance Neural tube Embryo Cell Biology Biological Sciences apico-basal polarity biology.organism_classification zebrafish Protein subcellular localization prediction 3. Good health Transport protein Cell biology Multicellular organism Protein Transport 030104 developmental biology medicine.anatomical_structure Signal transduction Developmental Biology Signal Transduction |
| Zdroj: | Developmental Cell Developmental cell, vol 36, iss 1 Buckley, C, Moore, R, Reade, A, Goldberg, A R, Weiner, O D & Clarke, J 2016, ' Reversible Optogenetic Control of Subcellular Protein Localization in a Live Vertebrate Embryo ', Developmental Cell, vol. 36, no. 1, pp. 117–126 . https://doi.org/10.1016/j.devcel.2015.12.011 Buckley, CE; Moore, RE; Reade, A; Goldberg, AR; Weiner, OD; & Clarke, JDW. (2016). Reversible Optogenetic Control of Subcellular Protein Localization in a Live Vertebrate Embryo. Developmental Cell, 36(1), 117-126. doi: 10.1016/j.devcel.2015.12.011. UCSF: Retrieved from: http://www.escholarship.org/uc/item/1qw221pt |
| ISSN: | 1878-1551 1534-5807 |
| DOI: | 10.1016/j.devcel.2015.12.011 |
| Popis: | Summary We demonstrate the utility of the phytochrome system to rapidly and reversibly recruit proteins to specific subcellular regions within specific cells in a living vertebrate embryo. Light-induced heterodimerization using the phytochrome system has previously been used as a powerful tool to dissect signaling pathways for single cells in culture but has not previously been used to reversibly manipulate the precise subcellular location of proteins in multicellular organisms. Here we report the experimental conditions necessary to use this system to manipulate proteins in vivo. As proof of principle, we demonstrate that we can manipulate the localization of the apical polarity protein Pard3 with high temporal and spatial precision in both the neural tube and the embryo’s enveloping layer epithelium. Our optimizations of optogenetic component expression and chromophore purification and delivery should significantly lower the barrier for establishing this powerful optogenetic system in other multicellular organisms. Graphical Abstract Highlights • The phytochrome system has been optimized for use within multicellular organisms • Protein recruitment can be tightly controlled to a specific subcellular region • Protein recruitment occurs with high binding and reversal kinetics • The subcellular localization of the apical polarity protein Pard3 is manipulated The phytochrome system is a powerful tool that is used to manipulate protein localization. Here, Buckley et al. optimize the phytochrome system in live zebrafish embryos and demonstrate rapid and reversible protein recruitment to specific subcellular regions within specific cells, at a high spatial and temporal resolution. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|