title: Risk-managed production of bioactive recombinant proteins using a novel plant virus vector with a helper plant to complement viral systemic movement.
abstract: 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.
authors.affiliation: 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.
authors.forename: Noriho
authors.initials: N
authors.lastname: Fukuzawa
authors.forename: Takeaki
authors.initials: T
authors.lastname: Ishihara
authors.forename: Noriko
authors.initials: N
authors.lastname: Itchoda
authors.forename: Noriko
authors.initials: N
authors.lastname: Tabayashi
authors.forename: Chiwa
authors.initials: C
authors.lastname: Kataoka
authors.forename: Chikara
authors.initials: C
authors.lastname: Masuta
authors.forename: Takeshi
authors.initials: T
authors.lastname: Matsumura
mesh_terms: D018501:Antirheumatic Agents / Q000378:metabolism; D001172:Arthritis, Rheumatoid / Q000188:drug therapy; D056738:Cross Protection; D017799:Cucumovirus / Q000235:genetics / Q000378:metabolism*; D004147:Dioxins / Q000276:immunology*; D018506:Gene Expression Regulation, Plant; D005822:Genetic Vectors; D030821:Plants, Genetically Modified; D017472:Receptors, Interleukin-1 / Q000037:antagonists & inhibitors*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012306:Risk; D057127:Single-Chain Antibodies / Q000096:biosynthesis; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D019076:Transgenes
chemical_list: D018501:Antirheumatic Agents; D004147:Dioxins; D017472:Receptors, Interleukin-1; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies