CRISPR/Cas9-Mediated Gene Targeting during Embryogenesis in Swine

Ability to disrupt genes is essential in elucidating gene function. Unlike rodents or amphibians, it has been difficult to generate gene-targeted embryos in large animals. Therefore, studies of early embryo development have been hampered in large animals. A recent technology suggests that targeted mutations can be successfully introduced during embryogenesis, thus by-passing the need of breeding to produce gene-targeted embryos. This is particularly important in large animal models because of longer gestation period and higher animal cost. Here, we describe a specific approach to disrupt up to two genes simultaneously during embryogenesis using the CRISPR/Cas9 technology in swine. The approach can help understand the mechanism of zygotic genome activation in large animals.