Efficient Transgenesis in African Turquoise Killifish Using a Gibson Assembly-Based Tol2 Transposon System

The short-lived African killifish Nothobranchius furzeri is an attractive genetic model for vertebrate aging and regeneration studies. The utilization of genetically modified animals is a common strategy for unveiling molecular mechanisms responsible for a biological phenomenon. Here, we report a highly efficient protocol for generating transgenic African killifish using the Tol2 transposon system, which creates random insertions in the genome. Transgenic vectors carrying gene-expression cassettes of interest and an eye-specific marker for transgene identification can be quickly assembled through Gibson assembly. The development of this new pipeline will facilitate transgenic reporter assays and gene-expression-related manipulations in African killifish.