Meier AF Laimbacher AS 2020 HSV-1 amplicon vectors have been used as platforms for the generation of genetic vaccines against both DNA and RNA viruses. Mice vaccinated with such vectors encoding structural proteins from both foot-and-mouth disease virus and rotavirus were partially protected from challenge with wild-type virus (D'Antuono et al., Vaccine 28:7363-7372, 2010; Laimbacher et al., Mol Ther 20:1810-1820, 2012; Meier et al., Int J Mol Sci 18:431, 2017), indicating that HSV-1 amplicon vectors are attractive tools for the development of complex and safe genetic vaccines.This chapter describes the preparation and testing of HSV-1 amplicon vectors that encode individual or multiple viral structural proteins from a polycistronic transgene cassette. We further put particular emphasis on generating virus-like particles (VLPs) in vector-infected cells. Expression of viral genes is confirmed by Western blot and immune fluorescence analysis and generation of VLPs in vector-infected cells is demonstrated by electron microscopy. Furthermore, examples on how to analyze the immune response in a mouse model and possible challenge experiments are described. https://sonar.ch/global/documents/226372 eng info:eu-repo/semantics/altIdentifier/doi/10.1007/978-1-4939-9814-2_6 info:eu-repo/semantics/altIdentifier/pmid/31617175 Methods in molecular biology (Clifton, N.J.). - 2020 Genetic vaccine Helper virus-free HSV-1 amplicon vector Polycistronic transgene cassette VLPs Virus-like particles Animals Chlorocebus aethiops Genetic Vectors Herpesvirus 1, Human Humans Mice Transduction, Genetic Vero Cells Viral Structural Proteins Viral Vaccines HSV-1 Amplicon Vectors as Genetic Vaccines. http://purl.org/coar/resource_type/c_6501