%0 Journal Article
%T Insertion of foreign genes into the simian varicella virus genome by Tn7-mediated site-specific transposition.
%A Gray WL
%J J Virol Methods
%V 327
%N 0
%D 2024 Jun 6
%M 38583808
%F 2.623
%R 10.1016/j.jviromet.2024.114936
%X A Tn7-transposition approach was utilized for site-specific insertion of foreign genes into the genome of simian varicella virus (SVV), the causative agent of simian varicella in nonhuman primates. The severe acute respiratory syndrome coronavirus (SARS-CoV-2) nucleocapsid (N) gene and receptor binding domain (RBD) of the spike gene were inserted into the ORF 14 region of the SVV genome cloned into a bacterial artificial chromosome and then transfected into Vero cells to generate the infectious recombinant SVV (rSVV). The rSVV replicated efficiently in infected Vero cells and expressed the N and RBD antigens as indicated by immunoblot and immunofluorescence assays. Tn7-mediated transposition provides a rapid and efficient method for constructing rSVVs which may be evaluated as live-attenuated vaccines.