Division of Infectious Diseases and International Medicine, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA.
Microbiology, Immunology, and Cancer Biology Graduate Program, University of Minnesota, Minneapolis, MN 55455, USA.
STAR Protoc. 2020 Dec 9;1(3):100209. doi: 10.1016/j.xpro.2020.100209. eCollection 2020 Dec 18.
We describe the production of single-cycle (sc) and replication-competent recombinant vesicular stomatitis viruses (rcVSVs) displaying heterologous envelope glycoproteins (Envs) on their surface. We prepare scVSVs by transiently expressing HIV-1 Envs or SARS-CoV-2 spike followed by infection of the cells with scVSV particles, which do not carry the gene. To prepare rcVSVs, we replace the vsv-g with a specific -encoding gene, transfect cells with multiple plasmids for production of the genomic RNA and viral proteins, and rescue replication-competent viruses.
我们描述了生产单周期（sc）和具有复制能力的重组水疱性口炎病毒（rcVSV）的方法，这些病毒在其表面展示异源包膜糖蛋白（Env）。我们通过瞬时表达 HIV-1 Env 或 SARS-CoV-2 刺突蛋白，然后用不携带 基因的 scVSV 颗粒感染细胞来制备 scVSV。为了制备 rcVSV，我们用特定的 基因替换 vsv-g，用多个质粒转染细胞以产生基因组 RNA 和病毒蛋白，并拯救具有复制能力的病毒。
