title: Plant Expression of Trans-Encapsidated Viral Nanoparticle Vaccines with Animal RNA Replicons.
abstract: In this protocol, we outline how to produce a live viral nanoparticle vaccine in a biosafety level 1 (BSL1) environment. An animal viral vector RNA encapsidated with tobacco mosaic virus (TMV) coat protein can be fully assembled in planta. Agrobacterium cultures containing each component are inoculated together into tobacco leaves and the self-assembled hybrid nanoparticle vaccine is harvested 4 days later and purified with a simple PEG precipitation. The viral RNA delivery vector is derived from the BSL1 insect virus, Flock House virus (FHV), and replicates in human and animal cells but does not spread systemically. A polyethylene glycol purification protocol is also provided to collect and purify these vaccines for immunological tests.
authors.affiliation: Institute of Biomedical Studies, Baylor University, Waco, TX, USA.
authors.forename: Yiyang
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: College of Pharmacy, Touro University California, Vallejo, CA, USA.
authors.forename: Alison A
authors.initials: AA
authors.lastname: McCormick
authors.affiliation: Institute of Biomedical Studies, Baylor University, Waco, TX, USA. Chris_Kearney@baylor.edu.|Department of Biology, Baylor University, Waco, TX, USA. Chris_Kearney@baylor.edu.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Kearney
mesh_terms: D000818:Animals; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D053758:Nanoparticles / Q000821:virology*; D012367:RNA, Viral / Q000235:genetics*; D012093:Replicon / Q000235:genetics*; D014026:Nicotiana / Q000821:virology*; D014027:Tobacco Mosaic Virus / Q000276:immunology; D014765:Viral Vaccines / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics
chemical_list: D012367:RNA, Viral; D014765:Viral Vaccines
keywords: Agroinoculation; Polyethylene glycol purification; Trans-encapsidation; Viral vaccine