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A removable virus vector suitable for plant genome editing.

Tetsuya Chujo1, Manabu Yoshikawa1, Hirotaka Ariga1

  • 1Plant and Microbial Research Unit, Division of Plant and Microbial Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan.

The Plant Journal : for Cell and Molecular Biology
|April 22, 2017
PubMed
Summary

Researchers developed a removable RNA virus vector for plant genome editing. This vector is eliminated during plant regeneration, enabling virus-free, edited plants without transgene integration.

Keywords:
genome editingmiR398technical advancetomato mosaic virusvirus vector

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Area of Science:

  • Plant Biotechnology
  • Molecular Biology
  • Virology

Background:

  • Plant genome editing relies on sequence-specific nucleases (SSNs).
  • RNA virus vectors offer transgene integration-free gene editing but pose removal challenges.
  • Existing methods lack effective antiviral strategies for plant virus vector clearance.

Purpose of the Study:

  • To engineer a removable RNA virus vector for plant genome editing.
  • To achieve transgene integration-free targeted mutagenesis in plants.
  • To develop a method for efficient virus vector elimination post-editing.

Main Methods:

  • Developed a novel RNA virus vector incorporating a target site for tobacco microRNA398 (miR398).
  • Utilized shoot regeneration to induce miR398 expression for viral RNA elimination.
  • Expressed SSNs via the virus vector for targeted mutagenesis in inoculated leaves.

Main Results:

  • The modified virus vector remained functional in inoculated leaves prior to miR398 induction.
  • Regenerating shoots triggered miR398 expression, leading to effective viral RNA clearance.
  • Successfully regenerated virus-free, genome-edited plants from tissue culture.

Conclusions:

  • A removable RNA virus vector system was successfully established for plant genome editing.
  • This approach enables transgene integration-free mutagenesis and efficient virus vector removal.
  • The method facilitates the regeneration of virus-free, genome-edited plants.