The maize was modified through a site-specific transformation protocol for insect resistance and herbicide tolerance. For resistance to Western corn rootworm (Diabrotica virgifera), the maize expresses Ophioderma pendulum insecticidal protein IPD079Ea, which has a pore-forming mode of action against feeding larvae. The protein binds receptors in the insect's midgut which are different from the receptors that Bt toxins interact with, allowing the maize to overcome Bt-resistance in Coleoptera pests. For tolerance to glufosinate, the maize expresses Streptomyces viridochromogenes phosphinothricin N-acetyltransferase, which inactivates the herbicidal compound through acetylation. In addition, the maize also contains an Escherichia coli phosphomannose isomerase cassette, which allows for modified plants to use mannose as a carbon source and thus is a selectable marker during transformation. The transformation protocol involved two steps to achieve a site specific integration into the maize genome. In the first step, four plasmids were introduced by microparticle bombardment, which introduced recombination sites at a specific location using transiently expressed CRISPR/Cas9. In the second step, Agrobacterium-mediated transformation was used to introduce the final gene cassettes through a recombination with sequences present in the first insertion site. More information regarding the two-step site-specific transformation is provided below.