Whole-genome Sequence Analysis to Confirm the Absence of Transgene in a Rice Line Made by Gene Targeting

DOI Web Site 参考文献22件
  • MANO Junichi
    Institute of Food Research, National Agriculture and Food Research Organization
  • TSUKAHARA Keita
    Institute of Food Research, National Agriculture and Food Research Organization Fasmac Co. Ltd.,
  • TAKASAKI Kazuto
    Fasmac Co. Ltd.,
  • FUTO Satoshi
    Fasmac Co. Ltd.,
  • NISHIZAWA-YOKOI Ayako
    Institute of Agrobiological Sciences, National Agriculture and Food Research Organization
  • TOKI Seiichi
    Institute of Agrobiological Sciences, National Agriculture and Food Research Organization Graduate School of Nanobioscience, Yokohama City University Kihara Institute for Biological Research, Yokohama City University Department of Plant Life Science, Faculty of Agriculture, Ryukoku University
  • TAKABATAKE Reona
    Institute of Food Research, National Agriculture and Food Research Organization
  • KITTA Kazumi
    Institute of Food Research, National Agriculture and Food Research Organization

説明

<p>The recent development of homologous recombination-mediated gene targeting (GT) techniques has made it easy to modify nucleotide sequences in plant genomes. Backcross breeding following GT provides transgene-free plant lines with high probability. However, owing to the possibility of unintentional transgene introduction during genetic transformation, analytical methods may be necessary to reliably and comprehensively detect transgenes remaining on a plant genome. Herein, we conducted an analysis based on whole-genome sequencing. We analyzed the GT rice DNA using a next-generation sequencer, and we performed data analysis to find the transgene integration on the genome. All results supported the absence of transgenes in GT rice. Our genome integrity evaluation method could also be applicable to plant lines produced using other genome-editing technologies such as CRISPR/Cas9.</p>

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