Figure 1.

Overview of Integration Specific Selection. A) The Synthetic Landing Pad (SLP) with synthetic guide RNA target sequence, the 3′ fragment of the hygromycin resistance gene (partial coding sequence and UTR), and a single loxP sequence. B) The donor homology with cargo transgene to be inserted, a second loxP sequence, and the 5′ fragment of the hygromycin resistance gene (promoter & partial coding sequence). C) Cas9 & guide expression plasmid is injected with donor homology. Cas9 targets and creates a double-strand break at the synthetic target location. D) Once the double-strand break is made, repair with the donor homology integrates the transgenic cargo, and restores the hygromycin gene, allowing selection to occur only upon integration.