Repositorium
Targeted deletion and inversion of tandemly arrayed genes in Arabidopsis thaliana using zinc finger nucleases
172
Journal Article / 2014
Qi, Yiping; Li, Xiaohong; Zhang, Yong; Starker, Colby G.; Baltes, Nicholas J.; Zhang, Feng; Sander, Jeffry D.; Reyon, Deepak; Joung, J. Keith; Voytas, Daniel F.
Tandemly arrayed genes (TAGs) or gene clusters are prevalent in higher eukaryotic genomes. For example, approximately 17% of genes are organized in tandem in the model plant Arabidopsis thaliana. The genetic redundancy created by TAGs presents a challenge for reverse genetics. As molecular scissors, engineered zinc finger nucleases (ZFNs) make DNA double-strand breaks in a sequence-specific manner. ZFNs thus provide a means to delete TAGs by creating two double-strand breaks in the gene cluster. Using engineered ZFNs, we successfully targeted seven genes from three TAGs on two Arabidopsis chromosomes, including the well-known RPP4 gene cluster, which contains eight resistance (R) genes. The resulting gene cluster deletions ranged from a few kb to 55 kb with frequencies approximating 1% in somatic cells. We also obtained large chromosomal deletions of ~9 Mb at approximately one tenth the frequency, and gene cluster inversions and duplications also were achieved. This study demonstrates the ability to use sequence-specific nucleases in plants to make targeted chromosome rearrangements and create novel chimeric genes for reverse genetics and biotechnology.
Techniques
ID | Corresponding Author Country |
Plant Species | GE Technique Sequence Identifier |
Trait Type of Alteration |
Progress in Research Key Topic |
---|---|---|---|---|---|
371 |
Voytas, Daniel F. USA |
Arabidopsis thaliana |
Zinc-finger nucleases ASK8 |
No information SDN1 |
Basic research Basic research |
372 |
Voytas, Daniel F. USA |
Arabidopsis thaliana |
Zinc-finger nucleases RLK |
No information SDN1 |
Basic research Basic research |