PUBLICATION
Reversible Optogenetic Control of Subcellular Protein Localization in a Live Vertebrate Embryo
- Authors
- Buckley, C.E., Moore, R.E., Reade, A., Goldberg, A.R., Weiner, O.D., Clarke, J.D.
- ID
- ZDB-PUB-160115-1
- Date
- 2016
- Source
- Developmental Cell 36: 117-126 (Journal)
- Registered Authors
- Clarke, Jon, Reade, Anna
- Keywords
- Pard3, apico-basal polarity, asymmetric inheritance, optogenetics, phytochrome, zebrafish
- MeSH Terms
-
- Animals
- Light
- Optogenetics*
- Protein Transport
- Signal Transduction/physiology*
- Zebrafish/embryology
- Zebrafish/genetics
- Zebrafish/metabolism*
- PubMed
- 26766447 Full text @ Dev. Cell
Citation
Buckley, C.E., Moore, R.E., Reade, A., Goldberg, A.R., Weiner, O.D., Clarke, J.D. (2016) Reversible Optogenetic Control of Subcellular Protein Localization in a Live Vertebrate Embryo. Developmental Cell. 36:117-126.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping