Microinjection is predominantly used to produce genetically modified fish. However, there are certain difficulties involved in some fish species. In this study, we tested an alternative method to produce genetically modified zebrafish by delivering DNA and other materials into embryos by electroporation. We optimized the electroporation conditions of a square wave electroporation system that work efficiently for the introduction of plasmid DNA, recombinant Cas9 nuclease and synthetic dual guide RNAs. Transgenic expression was observed in a wide range of tissues, which is comparable with those obtained by microinjection. We further determined that efficient gene delivery can be achieved during the cleavage stage. This study describes detailed electroporation parameters for gene delivery with high efficiency and low toxicity, providing a novel method to generate transgenic lines and genome editing.

显微注射主要用于生产转基因鱼。然而，某些鱼类存在一定的困难。在这项研究中，我们测试了另一种生产转基因斑马鱼的方法，即通过电穿孔将 DNA 和其他材料传递到胚胎中。我们优化了方波电穿孔系统的电穿孔条件，该系统可有效引入质粒 DNA、重组 Cas9 核酸酶和合成双引导 RNA。在广泛的组织中观察到转基因表达，这与通过显微注射获得的结果相当。我们进一步确定在切割阶段可以实现有效的基因递送。这项研究描述了高效、低毒的基因递送的详细电穿孔参数，为生成转基因株系和基因组编辑提供了一种新方法。