Vesicular stomatitis virus (VSV)-based vaccination has shown protective efficacy against filovirus infection. Following the approval of a VSV-based vaccine against Ebola virus, there have been efforts toward applying the same platform for other filoviruses, including Marburg virus (MARV) and Sudan virus. Because these vaccines express filovirus glycoproteins, they are also a valuable tool to study filovirus entry under biosafety level 2 conditions. In the protocol described below, we outline how to genetically manipulate a full-length VSV vector by removing the native VSV glycoprotein and replacing it with the surface-expressed MARV glycoprotein. In addition, we describe the recovery procedure of these recombinant, full-length VSVs and detail the necessary steps of virus propagation.
Keywords: ERVEBO; VSV; VSV-EBOV; VSV-MARV; Vaccine; Virus entry.
© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.