PUBLICATION
zGrad is a nanobody-based degron system that inactivates proteins in zebrafish
- Authors
- Yamaguchi, N., Colak-Champollion, T., Knaut, H.
- ID
- ZDB-PUB-190209-17
- Date
- 2019
- Source
- eLIFE 8: (Journal)
- Registered Authors
- Knaut, Holger
- Keywords
- cell biology, developmental biology, zebrafish
- MeSH Terms
-
- Nuclear Proteins/genetics*
- Proteolysis*
- Drosophila melanogaster/genetics
- Green Fluorescent Proteins/genetics
- Green Fluorescent Proteins/isolation & purification
- Cytosol/metabolism
- Zebrafish/genetics*
- Animals
- Drosophila/genetics
- Animals, Genetically Modified/genetics
- Single-Domain Antibodies/genetics*
- Single-Domain Antibodies/immunology
- PubMed
- 30735119 Full text @ Elife
Citation
Yamaguchi, N., Colak-Champollion, T., Knaut, H. (2019) zGrad is a nanobody-based degron system that inactivates proteins in zebrafish. eLIFE. 8:.
Abstract
The analysis of protein function is essential to modern biology. While protein function has mostly been studied through gene or RNA interference, more recent approaches to degrade proteins directly have been developed. Here, we adapted the anti-GFP nanobody-based system deGradFP from flies to zebrafish. We named this system zGrad and show that zGrad efficiently degrades transmembrane, cytosolic and nuclear GFP-tagged proteins in zebrafish in an inducible and reversible manner. Using tissue-specific and inducible promoters in combination with functional GFP-fusion proteins, we demonstrate that zGrad can inactivate transmembrane, cytosolic and nuclear proteins globally, locally and temporally with different consequences. Global protein depletion results in phenotypes similar to loss of gene activity while local and temporal protein inactivation yields more restricted and novel phenotypes. Thus, zGrad is a versatile tool to study the spatial and temporal requirement of proteins in zebrafish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping