Targeted Deletion and Inversion of Tandemly Arrayed Genes in Arabidopsis thaliana Using Zinc Finger Nucleases

Details

• Alternative Title:
  G3 (Bethesda)
• Personal Author:
  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.
• Description:
  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.
• Subjects:
  Arabidopsis Base Sequence Chromosome Deletion Deletion Endodeoxyribonucleases Genes, Plant Gene Targeting Inversion Investigations Molecular Sequence Data Multigene Family Mutagenesis, Site-Directed Plants, Genetically Modified Tandemly Arrayed Genes (TAGs) Tandem Repeat Sequences Zinc Finger Nuclease (ZFN) Zinc Fingers

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• Source:
  G3 (Bethesda). 2013; 3(10):1707-1715.
• Pubmed ID:
  23979943
• Pubmed Central ID:
  PMC3789795
• Document Type:
  Journal Article
• Funding:
  DP1 GM105378/DP/NCCDPHP CDC HHS/United States ; DP1 GM105378/GM/NIGMS NIH HHS/United States ; R01 GM088040/GM/NIGMS NIH HHS/United States ; R01 GM088040/GM/NIGMS NIH HHS/United States ; R01 GM098861/GM/NIGMS NIH HHS/United States
• Volume:
  3
• Issue:
  10
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:e7668813fd9f2808aeefc408ab8817f308b97176a9a60c795d6adf2dba6d9392
• Download URL:
  https://stacks.cdc.gov/view/cdc/22559/cdc_22559_DS1.pdf
• File Type:
  [PDF - 1.79 MB ]