Date: 18 Jan 2013

Gene Replacement

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Gene-targeting (GT) is a powerful tool for functional analysis of genes and for gene replacement in biotechnological applications, but unfortunately it is very inefficient in plants, because integration of foreign DNA mainly occurs via non-homologous recombination. Inactivation of the most conserved pathway of non-homologous recombination called non-homologous end-joining (NHEJ) did not result in much higher GT frequency. However, introduction of a double strand break (DSB) at the intended site of recombination significantly stimulated DNA recombination at this DSB site, which may lead to replacement of native DNA sequences with foreign DNA sequences. Artificial zinc finger nucleases (ZFNs) are nowadays the enzymes of choice to induce DSBs at specifically selected sites in the genome. Such DSB may lead to targeted mutagenesis through inaccurate repair via end-joining or to GT through repair via homologous recombination (HR).