Figure 8.

Usage of ZFN technology to bring the site-specific recombination tool to a locus of interest for future use. As described in Figure 7, a DSB can be induced at an allele in a locus of interest by a custom-designed ZFN and repaired through HR pathway. This in turn can introduce a GOI-lox-SMG-lox or GOI-attB construct, for example, into the locus. The SMG can be deleted through recombinase-mediated site-specific deletion. In the second case, the attB site can be used for transgene-stacking at this locus of interest. Abbreviations. GOI: gene of interest, SMG: selectable marker gene, lox: loxP site of Cre-lox site-specific recombination system.