ZFIN ID: ZDB-PUB-190209-17
zGrad is a nanobody-based degron system that inactivates proteins in zebrafish
Yamaguchi, N., Colak-Champollion, T., Knaut, H.
Date: 2019
Source: eLIFE   8: (Journal)
Registered Authors: Knaut, Holger
Keywords: cell biology, developmental biology, zebrafish
MeSH Terms:
  • Animals
  • Animals, Genetically Modified/genetics
  • Cytosol/metabolism
  • Drosophila/genetics
  • Drosophila melanogaster/genetics
  • Green Fluorescent Proteins/genetics
  • Green Fluorescent Proteins/isolation & purification
  • Nuclear Proteins/genetics*
  • Proteolysis*
  • Single-Domain Antibodies/genetics*
  • Single-Domain Antibodies/immunology
  • Zebrafish/genetics*
PubMed: 30735119 Full text @ Elife
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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.
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