• Open Access

Risk-managed production of bioactive recombinant proteins using a novel plant virus vector with a helper plant to complement viral systemic movement

Authors

  • Noriho Fukuzawa,

    1. Plant Molecular Technology Research Group, Research Institute of Genome-based Biofactory, National Institute of Advanced Industrial Science and Technology (AIST), Tsukisamuhigashi Toyohira-Ku, Sapporo, Japan
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  • Takeaki Ishihara,

    1. Plant Breeding & Production Division, Agricultural Research Institute, HOKUREN Federation of Agricultural Cooperatives, Naganuma, Hokkaido, Japan
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  • Noriko Itchoda,

    1. Plant Breeding & Production Division, Agricultural Research Institute, HOKUREN Federation of Agricultural Cooperatives, Naganuma, Hokkaido, Japan
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  • Noriko Tabayashi,

    1. Plant Molecular Technology Research Group, Research Institute of Genome-based Biofactory, National Institute of Advanced Industrial Science and Technology (AIST), Tsukisamuhigashi Toyohira-Ku, Sapporo, Japan
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  • Chiwa Kataoka,

    1. Carbuncle BioScienTechLLC, Tanida, Okukaiinji, Nagaokakyo, Kyotohu, Japan
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  • Chikara Masuta,

    Corresponding author
    1. Graduate School of Agriculture, Hokkaido University, Sapporo, Japan
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  • Takeshi Matsumura

    1. Plant Molecular Technology Research Group, Research Institute of Genome-based Biofactory, National Institute of Advanced Industrial Science and Technology (AIST), Tsukisamuhigashi Toyohira-Ku, Sapporo, Japan
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(Tel +81 11 706 2807; fax +81 11 706 2483; email masuta@res.agr.hokudai.ac.jp)

Summary

A plant viral vector has the potential to efficiently produce recombinant proteins at a low cost in a short period. Although recombinant proteins can be also produced by transgenic plants, a plant viral vector, if available, may be more convenient when urgent scale-up in production is needed. However, it is difficult to use a viral vector in open fields because of the risk of escape to the environment. In this study, we constructed a novel viral vector system using a movement-defective Cucumber mosaic virus (CMV) vector, which is theoretically localized in the inoculated cells but infects systemically only with the aid of the transgenic helper plant that complements viral movement, diminishing the risk of viral proliferation. Interestingly, the helper plant systemically infected with the vector gave strong cross-protection against challenge inoculation with wild-type CMVs. Using CMV strains belonging to two discrete CMV groups (subgroups I and II), we also improved the system to prevent recombination between the vector and the transgene transcript in the helper plant. We here demonstrate the expression of an anti-dioxin single chain variable fragment (DxscFv) and interleukin-1 receptor antagonist (IL1-Ra) in Nicotiana benthamiana by this viral vector confinement system, which is applicable for many useful high-quality recombinant proteins.

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