Gene targeting by homologous recombination is a powerful method to manipulate the genome precisely and could be exploited to correct genetic defects. Zinc finger nucleases are designed proteins that fuse a zinc finger DNA binding domain to the nuclease domain from the FokI restriction endonuclease. Zinc finger nucleases were generated that stimulated gene targeting from half-site sequences from the human β-globin gene and the human common γ-chain gene. Zinc finger nucleases were also generated that stimulated gene targeting at full sites from the green fluorescent protein gene and the human CD8α gene. This work built on the prior zinc finger design work of others and in targeting these four genes had a 100% success rate at designing nucleases to the consensus half-site 5′-GNNGNNGNN-3′ and the consensus full site 5′-NNCNNCNNCNNNNNNGNNGNNGNN-3′, suggesting that zinc finger nucleases can be empirically designed to stimulate gene targeting in a large portion of the mammalian genome. Copyright © The American Society of the Gene Therapy.
Porteus, M. H. (2006). Mammalian gene targeting with designed zinc finger nucleases. Molecular Therapy, 13(2), 438–446. https://doi.org/10.1016/j.ymthe.2005.08.003