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Reversible Optogenetic Control of Subcellular Protein Localization in a Live Vertebrate Embryo.

Published version
Peer-reviewed

Type

Article

Change log

Authors

Buckley, Clare E 
Moore, Rachel E 
Reade, Anna 
Goldberg, Anna R 
Weiner, Orion D 

Abstract

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.

Description

Keywords

Pard3, apico-basal polarity, asymmetric inheritance, optogenetics, phytochrome, zebrafish, Animals, Light, Optogenetics, Protein Transport, Signal Transduction, Zebrafish

Journal Title

Dev Cell

Conference Name

Journal ISSN

1534-5807
1878-1551

Volume Title

36

Publisher

Elsevier BV