Efficient Gene Targeting with Zinc-finger Nucleases
Zinc-finger nucleases (ZFNs) are targetable DNA cleavage reagents that have been adopted as gene targeting tools. ZFN-induced double-strand breaks are subject to cellular DNA repair processes that lead to both targeted mutagenesis and targeted gene replacement at remarkably high frequencies. This talk will briefly review the history of ZFN development and summarize applications that have been made to genome alterations in many different organisms. Most of the work in my lab has been done with the fruit fly, Drosophila melanogaster, where we have explored mechanisms of repair of ZFN-induced breaks and attempted to optimize aspects of the gene targeting procedure. Recently we have focused our attention on properties of the donor DNA that affect the efficiency of gene replacement. Other issues that impact the utility of ZFNs will also be addressed.