PUBLICATION

A GCSFR/CSF3R zebrafish mutant models the persistent basal neutrophil deficiency of severe congenital neutropenia

Authors
Pazhakh, V., Clark, S., Keightley, M.C., Lieschke, G.J.
ID
ZDB-PUB-170311-3
Date
2017
Source
Scientific Reports   7: 44455 (Journal)
Registered Authors
Keightley, M. Cristina, Lieschke, Graham J., Pazhakh, Vahid
Keywords
Development, Mechanisms of disease, Myelopoiesis
MeSH Terms
  • Base Sequence
  • Leukocyte Count
  • Haploinsufficiency
  • Heterozygote
  • Gene Editing/methods*
  • Animals
  • Zebrafish
  • Morpholinos/genetics
  • Morpholinos/metabolism
  • Neutrophils/immunology
  • Neutrophils/pathology*
  • Granulocyte Colony-Stimulating Factor/genetics*
  • Granulocyte Colony-Stimulating Factor/immunology
  • Phenotype
  • Disease Models, Animal
  • CRISPR-Cas Systems
  • Gene Expression
  • Kidney/immunology
  • Kidney/pathology*
  • Humans
  • Receptors, Colony-Stimulating Factor/antagonists & inhibitors
  • Receptors, Colony-Stimulating Factor/deficiency
  • Receptors, Colony-Stimulating Factor/genetics*
  • Receptors, Colony-Stimulating Factor/immunology
  • Exons
  • Embryo, Nonmammalian
  • Neutropenia/congenital*
  • Neutropenia/genetics
  • Neutropenia/immunology
  • Neutropenia/pathology
(all 30)
PubMed
28281657 Full text @ Sci. Rep.
Abstract
Granulocyte colony-stimulating factor (GCSF) and its receptor (GCSFR), also known as CSF3 and CSF3R, are required to maintain normal neutrophil numbers during basal and emergency granulopoiesis in humans, mice and zebrafish. Previous studies identified two zebrafish CSF3 ligands and a single CSF3 receptor. Transient antisense morpholino oligonucleotide knockdown of both these ligands and receptor reduces neutrophil numbers in zebrafish embryos, a technique widely used to evaluate neutrophil contributions to models of infection, inflammation and regeneration. We created an allelic series of zebrafish csf3r mutants by CRISPR/Cas9 mutagenesis targeting csf3r exon 2. Biallelic csf3r mutant embryos are viable and have normal early survival, despite a substantial reduction of their neutrophil population size, and normal macrophage abundance. Heterozygotes have a haploinsufficiency phenotype with an intermediate reduction in neutrophil numbers. csf3r mutants are viable as adults, with a 50% reduction in tissue neutrophil density and a substantial reduction in the number of myeloid cells in the kidney marrow. These csf3r mutants are a new animal model of human CSF3R-dependent congenital neutropenia. Furthermore, they will be valuable for studying the impact of neutrophil loss in the context of other zebrafish disease models by providing a genetically stable, persistent, reproducible neutrophil deficiency state throughout life.
Genes / Markers
Figures
Figure Gallery (3 images)
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Expression
Phenotype
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
gl28TgTransgenic Insertion
    gl31
      Small Deletion
      gl32
        Indel
        gl33
          Small Deletion
          gl34
            Small Deletion
            i114TgTransgenic Insertion
              1 - 6 of 6
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              Human Disease / Model
              Sequence Targeting Reagents
              Target Reagent Reagent Type
              csf3rCRISPR1-csf3rCRISPR
              csf3rCRISPR2-csf3rCRISPR
              csf3rCRISPR3-csf3rCRISPR
              1 - 3 of 3
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              Fish
              Antibodies
              No data available
              Orthology
              No data available
              Engineered Foreign Genes
              Marker Marker Type Name
              GFPEFGGFP
              KALTA4EFGKALTA4
              1 - 2 of 2
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              Mapping
              No data available