title: Targeted deletion and inversion of tandemly arrayed genes in Arabidopsis thaliana using zinc finger nucleases.
abstract: 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.
authors: Department of Genetics, Cell Biology & Development and Center for Genome Engineering, University of Minnesota, Minneapolis, Minnesota 55455.
authors: Yiping
authors: Y
authors: Qi
authors: Xiaohong
authors: X
authors: Li
authors: Yong
authors: Y
authors: Zhang
authors: Colby G
authors: CG
authors: Starker
authors: Nicholas J
authors: NJ
authors: Baltes
authors: Feng
authors: F
authors: Zhang
authors: Jeffry D
authors: JD
authors: Sander
authors: Deepak
authors: D
authors: Reyon
authors: J Keith
authors: JK
authors: Joung
authors: Daniel F
authors: DF
authors: Voytas
mesh_terms: D017360:Arabidopsis / Q000235:genetics*; D001483:Base Sequence; D002872:Chromosome Deletion; D004706:Endodeoxyribonucleases / Q000737:chemistry / Q000235:genetics*; D018390:Gene Targeting / Q000379:methods*; D017343:Genes, Plant; D008969:Molecular Sequence Data; D005810:Multigene Family; D016297:Mutagenesis, Site-Directed / Q000379:methods; D030821:Plants, Genetically Modified / Q000235:genetics; D020080:Tandem Repeat Sequences; D016335:Zinc Fingers
chemical_list: D004706:Endodeoxyribonucleases
keywords: Arabidopsis; deletion; inversion; tandemly arrayed genes (TAGs); zinc finger nuclease (ZFN)
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