A removable virus vector suitable for plant genome editing |
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| Authors: | Tetsuya Chujo Manabu Yoshikawa Hirotaka Ariga Masaki Endo Seiichi Toki Kazuhiro Ishibashi |
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| Institution: | 1. Plant and Microbial Research Unit, Division of Plant and Microbial Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan;2. Plant Genome Engineering Research Unit, Division of Applied Genetics, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan;3. Graduate School of Nanobioscience, Yokohama City University, Yokohama, Kanagawa, Japan;4. Kihara Institute for Biological Research, Yokohama City University, Yokohama, Kanagawa, Japan |
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| Abstract: | Plant genome editing is achieved by the expression of sequence‐specific nucleases (SSNs). RNA virus vector‐mediated expression of SSNs is a promising approach for transgene integration‐free targeted mutagenesis in plants. However, the removal of virus vectors from infected plants is challenging because no antiviral drugs are available against plant viruses. Here, we developed a removable RNA virus vector that carries the target site of tobacco microRNA398 (miR398) whose expression is induced during shoot regeneration. In the inoculated leaves in which expression of miR398 is not induced, insertion of the miR398 target site did not affect the practicability of the virus vector. When shoots were regenerated from the infected leaves, miR398 was expressed and viral RNA was eliminated. The virus vector successfully expressed SSNs in inoculated leaves, from which virus‐free genome‐edited plants were regenerated via tissue culture. |
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| Keywords: | virus vector genome editing tomato mosaic virus miR398 technical advance |
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