Target-directed gene-editing approach for developing a new horticultural crop
Target mutagenesis of genes associated with preferred traits has been advancing continuously, and a precise technique applicable to genome modification of plants has been introduced recently. Adopted first for use with the animal genomes, target-gene editing using a nuclease has now been reported in some plant species including Arabidopsis, maize, tobacco, and other model systems. The genome-editing technology using the endonuclease depends on endogenously operating DNA-repair mechanisms such as non-homologous end joining (NHEJ) or homologous recombination (HR) and leads to insertion of a foreign DNA fragment or deletion of the target locus, which collectively allow changes in plant traits of interest. The nuclease is composed of two unique parts, a DNA-binding domain and a cleaving domain. A custom-designed TALEN was delivered into Petunia cells via an agrobacterium or PEG-mediated process, and then the engineered nuclease targeted genes conditioning crop traits of interest and modified them either constitutively or transiently. The transformant progenies will be screened visually or by PCR along with molecular confirmation analysis by using various tools. We have eventually extended the tools to other horticultural crop species and target genes, which is especially useful when a traditional breeding method is limited by lack of naturally occurring useful alleles or by some technical problems such as a reproduction barrier or gene fixation.
Lee, G.J., Kanth, B.K., Chung, S.J., Kim, S.J. and Bae, S. (2016). Target-directed gene-editing approach for developing a new horticultural crop. Acta Hortic. 1127, 289-294
mutagenesis, engineered nuclease, F3H, tissue culture, transient assay, Petunia