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

ABSTRACT: 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 f

SUBMITTER: Isiaku AI 

PROVIDER: S-EPMC8319549 | biostudies-literature | 2021 Jul

REPOSITORIES: biostudies-literature