Abstract
The induction of double-strand DNA breaks (DSBs) greatly increases rates of homologous recombination-mediated gene targeting or of gene mutation by error-prone non-homologous end joining. Both repair mechanisms would be important basis for molecular breeding to modify the targeted gene on the genome. However, these approaches depend on the capability to create a DSB on the specific genomic sequence of interest.
Zinc finger nucleases (ZFNs) have been recently developed to introduce a site-specific DSB. ZFNs function as dimers with each monomer composed of a non-specific cleavage domain from the Fok I endonuclease fused to a zinc-finger array engineered to bind a target DNA sequence of interest.
To demonstrate the ZFN approach to higher plants, especially rice, we developed systems for designing of ZFN, confirming digestion in in vitro and in vivo. We are on going to establish the system for induction of genome editing on the rice genome.
