Transient, flexible gene editing in zebrafish neutrophils and macrophages for determination of cell-autonomous functions

Abdulsalam I. Isiaku, Zuobing Zhang, Vahid Pazhakh, Harriet R. Manley, Ella R. Thompson, Lucy C. Fox, Satwica Yerneni, Piers Blombery, Graham J. Lieschke

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11 Citations (Scopus)


Zebrafish are an important model for studying phagocyte function, but rigorous experimental systems to distinguish whether phagocyte-dependent effects are neutrophil or macrophage specific have been lacking. We have developed and validated transgenic lines that enable superior demonstration of cell-autonomous neutrophil and macrophage genetic requirements. We coupled well-characterized neutrophil- and macrophage-specific Gal4 driver lines with UAS:Cas9 transgenes for selective expression of Cas9 in either neutrophils or macrophages. Efficient gene editing, confirmed by both Sanger and next-generation sequencing, occurred in both lineages following microinjection of efficacious synthetic guide RNAs into zebrafish embryos. In proof-of-principle experiments, we demonstrated molecular and/or functional evidence of on-target gene editing for several genes (mCherry, lamin B receptor, trim33) in either neutrophils or macrophages as intended. These new UAS:Cas9 tools provide an improved resource for assessing individual contributions of neutrophil- and macrophage-expressed genes to the many physiological processes and diseases modelled in zebrafish. Furthermore, this gene-editing functionality can be exploited in any cell lineage for which a lineage-specific Gal4 driver is available.

Original languageEnglish
Article numberdmm047431
Number of pages13
JournalDisease Models & Mechanisms
Issue number7
Publication statusPublished - Jul 2021


  • Cell autonomy
  • CRISPR-Cas9
  • Gene editing
  • Macrophages
  • Neutrophils
  • Zebrafish

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