Sandra Cristea, Yevgeniy Freyvert, Yolanda Santiago, Michael C. Holmes, Fyodor D. Urnov, Philip D. Gregory and Gregory J. Cost In vivo cleavage of transgene donors promotes nuclease-mediated targeted integration Biotechnology and Bioengineering 110
Targeted transgene integration is a critical tool for genome engineering. Unfortunately some organisms (including CHO and plant cells) are refractory to targeted transgenesis due to deficits in homology-based DNA repair. In this paper, Cristea and Freyvert, and coworkers develop a homology-independent targeted DNA integration technique based on zinc-finger nucleases that should allow for efficient targeted DNA integration in all organisms. Key to the method's success is linearization of the donor DNA in vivo by the zinc-finger nucleases.
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