The Structural Features of Thousands of T-DNA Insertion Sites Are Consistent with a Double-Strand Break Repair-Based Insertion Mechanism

Molecular Plant
Nils KleinboeltingBernd Weisshaar

Abstract

Transformation by Agrobacterium tumefaciens, an important tool in modern plant research, involves the integration of T-DNA initially present on a plasmid in agrobacteria into the genome of plant cells. The process of attachment of the agrobacteria to plant cells and the transport of T-DNA into the cell and further to the nucleus has been well described. However, the exact mechanism of integration into the host's DNA is still unclear, although several models have been proposed. During confirmation of T-DNA insertion alleles from the GABI-Kat collection of Arabidopsis thaliana mutants, we have generated about 34,000 sequences from the junctions between inserted T-DNA and adjacent genome regions. Here, we describe the evaluation of this dataset with regard to existing models for T-DNA integration. The results suggest that integration into the plant genome is mainly mediated by the endogenous plant DNA repair machinery. The observed integration events showed characteristics highly similar to those of repair sites of double-strand breaks with respect to microhomology and deletion sizes. In addition, we describe unexpected integration events, such as large deletions and inversions at the integration site that are relevant for correct...Continue Reading

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Citations

Nov 4, 2016·Nature Plants·Maartje van KregtenMarcel Tijsterman
Aug 31, 2017·Annual Review of Genetics·Stanton B Gelvin
Jan 16, 2019·Plant Molecular Biology·Keunsub LeeKan Wang
Jan 19, 2019·PLoS Genetics·Florian JupeJoseph R Ecker
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Aug 21, 2019·Frontiers in Genetics·Liang SunElison B Blancaflor
Oct 27, 2020·The New Phytologist·Ayako Nishizawa-YokoiStanton B Gelvin
Jun 8, 2021·Physiology and Molecular Biology of Plants : an International Journal of Functional Plant Biology·Agnieszka SkarzyńskaWojciech Pląder
Aug 28, 2021·International Journal of Molecular Sciences·Stanton B Gelvin
Oct 30, 2021·The Plant Journal : for Cell and Molecular Biology·Niels van TolMarcel Tijsterman

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