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
  • 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*
(all 12)
PubMed
30735119 Full text @ Elife
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
Marker Marker Type Name
cdh1GENEcadherin 1, type 1, E-cadherin (epithelial)
cryaaGENEcrystallin, alpha A
ctnna1GENEcatenin (cadherin-associated protein), alpha 1
cxcr4bGENEchemokine (C-X-C motif), receptor 4b
h2az2aGENEH2A.Z variant histone 2a
hsp70lGENEheat shock cognate 70-kd protein, like
myl7GENEmyosin, light chain 7, regulatory
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Figures
Figure Gallery (12 images) / 2
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Expression
Phenotype
No data available
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
ct3aGtTransgenic Insertion
sk76TgTransgenic Insertion
    sk84TgTransgenic Insertion
      sk87TgTransgenic Insertion
        sk92TgTransgenic Insertion
          sk93TgTransgenic Insertion
            sk94TgTransgenic Insertion
              sk95TgTransgenic Insertion
                sk96TgTransgenic Insertion
                  t26035
                    		Point Mutation
                    1 - 10 of 11
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                    Human Disease / Model
                    No data available
                    Sequence Targeting Reagents
                    No data available
                    Fish
                    No data available
                    Antibodies
                    No data available
                    Orthology
                    No data available
                    Engineered Foreign Genes
                    Marker Marker Type Name
                    mScarletEFGmScarlet
                    CeruleanEFGCerulean
                    CitrineEFGCitrine
                    DsRedEFGDsRed
                    EGFPEFGEGFP
                    GFPEFGGFP
                    mCherryEFGmCherry
                    mKate2EFGmKate2
                    TagBFPEFGTagBFP
                    TagRFPEFGTagRFP
                    1 - 10 of 11
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                    Mapping
                    No data available
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                    Citation
                    Yamaguchi, N., Colak-Champollion, T., Knaut, H. (2019) zGrad is a nanobody-based degron system that inactivates proteins in zebrafish. eLIFE. 8:.