Generation of cd63-deficient zebrafish to analyze the role of cd63 in viral infection

Jung, S., Kim, M.J., Sellaththurai, S., Kim, S., Lee, S., Lee, J.
Fish & shellfish immunology   111: 152-159 (Other)
Registered Authors
Kim, Myoung-Jin
CRISPR/Cas9 system, cd63 knockout, immune response, viral infection, zebrafish
MeSH Terms
  • Amino Acid Sequence
  • Animals
  • Fish Diseases/immunology*
  • Fish Proteins/chemistry
  • Fish Proteins/deficiency
  • Fish Proteins/genetics
  • Fish Proteins/immunology
  • Gene Expression Profiling/veterinary
  • Gene Expression Regulation/immunology*
  • Immunity/genetics*
  • Novirhabdovirus/physiology
  • Phylogeny
  • Rhabdoviridae Infections/immunology
  • Rhabdoviridae Infections/veterinary
  • Sequence Alignment/veterinary
  • Tetraspanin 30/chemistry
  • Tetraspanin 30/deficiency
  • Tetraspanin 30/genetics*
  • Tetraspanin 30/immunology*
  • Zebrafish/genetics*
  • Zebrafish/immunology*
33556552 Full text @ Fish Shellfish Immunol.
The tetraspanin superfamily proteins are transmembrane proteins identified in a diverse range of eukaryotic organisms. Tetraspanins are involved in a variety of essential biological functions, including cell differentiation, adhesion, migration, signal transduction, intracellular trafficking, and immune responses. For an infection to occur, viruses must interact with various cell surface components, including receptors and signaling molecules. Tetraspanin CD63 is involved in the organization of the cell membrane and trafficking of cellular transmembrane proteins that interact with many viruses. In this study, the cd63 gene was characterized by studying its expression and function in a zebrafish model. The functional domains and structural features of cd63, such as the Cys-Cys-Gly (CCG) motif in the large extracellular loop and cysteine residues, are conserved in zebrafish. We confirmed that cd63 was expressed in immune system organs, such as the axial vein and pronephric duct, during the embryonic development of zebrafish. To better understand the role of cd63 in the zebrafish immune system, we established cd63-deficient zebrafish lines using the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system. A 19 bp insertion mutation was generated in single guide RNA (sgRNA) target sequence of exon 3 of the cd63 gene, to create a pre-mature stop codon. We then analyzed the expression of cd63-related genes cxcr4a and cxcr4b in wild type (WT) and cd63-deficient zebrafish. We believe our study provides an important model that could be used to investigate the roles of cd63 in viral infection in vivo.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes