Journal
DEVELOPMENTAL CELL
Volume 36, Issue 1, Pages 117-126Publisher
CELL PRESS
DOI: 10.1016/j.devcel.2015.12.011
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Funding
- European Molecular Biology Organisation
- BBSRC
- Wellcome Trust
- AHA Established Investigator Award
- [GM084040]
- [GM096164]
- Biotechnology and Biological Sciences Research Council [BB/K000926/1] Funding Source: researchfish
- BBSRC [BB/K000926/1] Funding Source: UKRI
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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.
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