Rapidly generating homozygous mutate zebrafish in F0 generation by technical integration of CRISPR/Cas9 and gynogenesis

Gene editing technique has been widely applied for gene function characterization. However, such an approach is often time-consuming to obtain homozygous mutant. In this article, through timing the earlier injection of embryos, the gene editing efficiency is significantly improved, and together with the technique of the UV inactivated sperm and heat shock treatment for blocking the first cleavage, the gene-edited homozygous zebrafish are successfully obtained in F0 generation, rather than in the third or more generations as reported in the existing results. It is concluded that genetic purification of artificial gynogenesis can play critical role in more efficiently preparing the gene-edited homozygous individuals in fish.