PUBLICATION

Generation and Characterization of a CRISPR/Cas9 -Induced 3-mst Deficient Zebrafish

Authors
Katsouda, A., Peleli, M., Asimakopoulou, A., Papapetropoulos, A., Beis, D.
ID
ZDB-PUB-200225-2
Date
2020
Source
Biomolecules   10(2): (Journal)
Registered Authors
Beis, Dimitris
Keywords
3-mercaptopyruvate sulfurtransferase, hydrogen sulfide, reactive oxygen species, zebrafish
MeSH Terms
  • Animals
  • CRISPR-Cas Systems*
  • Hydrogen Sulfide/metabolism
  • Oxidative Stress*
  • Regeneration
  • Sulfurtransferases/genetics*
  • Sulfurtransferases/metabolism
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/physiology
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/metabolism
(all 12)
PubMed
32079278 Full text @ Biomolecules
Abstract
3-mercaptopyruvate sulfurtransferase (3-MST) is an enzyme capable of synthesizing hydrogen sulfide (H2S) and polysulfides. In spite of its ubiquitous presence in mammalian cells, very few studies have investigated its contribution to homeostasis and disease development, thus the role of 3-MST remains largely unexplored. Here, we present a clustered, regularly interspaced, short palindromic repeats (CRISPR)/CRISPR-associated protein-9 (Cas9) induced 3-mst mutant zebrafish line, which will allow the study of 3-MST's role in several biological processes. The 3-mst zebrafish orthologue was identified using a bioinformatic approach and verified by its ability to produce H2S in the presence of 3-mercaptopyruvate (3-MP). Its expression pattern was analyzed during zebrafish early development, indicating predominantly an expression in the heart and central nervous system. As expected, no detectable levels of 3-Mst protein were observed in homozygous mutant larvae. In line with this, H2S levels were reduced in 3-mst-/- zebrafish. Although the mutants showed no obvious morphological deficiencies, they exhibited increased lethality under oxidative stress conditions. The elevated levels of reactive oxygen species, detected following 3-mst deletion, are likely to drive this phenotype. In line with the increased ROS, we observed accelerated fin regenerative capacity in 3-mst deficient zebrafish. Overall, we provide evidence for the expression of 3-mst in zebrafish, confirm its important role in redox homeostasis and indicate the enzyme's possible involvement in the regeneration processes.
Genes / Markers
Marker Marker Type Name
mpstGENEmercaptopyruvate sulfurtransferase
1 - 1 of 1
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Figures
Figure Gallery (6 images)
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Expression
Phenotype
Fish Conditions Stage Phenotype Figure
mpstaa102/aa102standard conditionsDay 4
mpstaa102/aa102standard conditionsDay 4
mpstaa102/aa102standard conditionsDay 4
mpstaa102/aa102chemical treatment by environment: paraquatDay 6
mpstaa102/aa102chemical treatment by environment: paraquatDay 6
mpstaa102/aa102chemical treatment by environment: paraquatDay 6
mpstaa102/aa102amputation: caudal finAdult
1 - 7 of 7
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Mutations / Transgenics
Allele Construct Type Affected Genomic Region
aa102
    		Insertion
    1 - 1 of 1
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    Human Disease / Model
    No data available
    Sequence Targeting Reagents
    Target Reagent Reagent Type
    mpstCRISPR1-mpstCRISPR
    1 - 1 of 1
    Show
    Fish
    Fish
    AB
    mpstaa102/aa102
    1 - 2 of 2
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    Antibodies
    Name Type Antigen Genes Isotypes Host Organism
    Ab1-MPSTpolyclonal
      		IgGRabbit
      1 - 1 of 1
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      Orthology
      Gene Orthology
      mpst
      1 - 1 of 1
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      Engineered Foreign Genes
      No data available
      Mapping
      No data available
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      Citation
      Katsouda, A., Peleli, M., Asimakopoulou, A., Papapetropoulos, A., Beis, D. (2020) Generation and Characterization of a CRISPR/Cas9 -Induced 3-mst Deficient Zebrafish. Biomolecules. 10(2):.