PUBLICATION
An in vivo translation-reporter system for the study of protein synthesis in zebrafish embryos
- Authors
- Palha, I.G., Anselme, I., Schneider-Maunoury, S., Giudicelli, F.
- ID
- ZDB-PUB-181109-1
- Date
- 2018
- Source
- Biology Open 7(12): (Journal)
- Registered Authors
- Anselme, Isabelle, Schneider-Maunoury, Sylvie
- Keywords
- Local translation, Neuron, SPoT, TimeStamp, Zebrafish
- MeSH Terms
- none
- PubMed
- 30404898 Full text @ Biol. Open
Citation
Palha, I.G., Anselme, I., Schneider-Maunoury, S., Giudicelli, F. (2018) An in vivo translation-reporter system for the study of protein synthesis in zebrafish embryos. Biology Open. 7(12):.
Abstract
Control of gene expression at the translation level is increasingly regarded as a key feature in many biological processes. Simple, inexpensive, and reliable procedures to visualise sites of protein production are required to allow observation of the spatiotemporal patterns of mRNA translation at subcellular resolution. We present a method, named SPoT (for Subcellular Patterns of Translation), developed upon the original TimeStamp technique (Lin et al., 2008), consisting in the expression of a fluorescent protein fused to a tagged, self-cleavable protease domain. Addition of a cell-permeable protease inhibitor instantly stabilizes newly produced, tagged protein allowing to distinguish recently synthesized protein from preexisting one. After a brief protease inhibitor treatment, the ratio of tagged vs non-tagged forms is highest at sites where proteins are the most recent, i.e. sites of synthesis. Therefore, by comparing tagged and non-tagged protein it is possible to spotlight sites of translation. By specifically expressing the SPoT cassette in neurons of transgenic zebrafish embryos, we reveal sites of neuronal protein synthesis in diverse cellular compartments during early development.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping