期刊
G3-GENES GENOMES GENETICS
卷 3, 期 10, 页码 1707-1715出版社
GENETICS SOCIETY AMERICA
DOI: 10.1534/g3.113.006270
关键词
zinc finger nuclease (ZFN); tandemly arrayed genes (TAGs); deletion; inversion; Arabidopsis
资金
- National Science Foundation [MCB 0209818, DBI 0923827]
- National Institutes of Health (NIH)
- Chief Science Officer of Cellectis Plant Sciences
- [DP1 GM105378]
- [R01 GM088040]
- Direct For Biological Sciences
- Division Of Integrative Organismal Systems [0923827] Funding Source: National Science Foundation
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 similar to 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.
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