Protocol for constructing and characterizing recombinant vectored vaccines for rabies virus
Year: 2024
Type: article
Abstract: Recombinant vectored vaccines provide economical, rapidly engineered, and tailored immunization strategies. Here, we present a protocol for constructing and characterizing recombinant vectored vaccines for rabies virus (RV). We describe steps for generating replication-competent vesicular stomatitis virus (rcVSV) by replacing vesicular stomatitis virus (VSV) glycoprotein (G) with RV G gene and for recovering rcVSV by co-transfecting baby hamster kidney (BHK-21) cells with the antigenomic VSV cDNA plasmid (pVSV). We then detail procedures for the functional, structural, and molecular characterization of generated rcVSV-RV-Gs. • Steps to display heterologous viral glycoproteins on recombinant VSVs • Procedures for rescuing recombinant VSV in mammalian cells • Instructions to evaluate the replication fitness of rcVSV-RV-G • Steps for assessing the expression of RV-G and VSV-M proteins by western blot Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Recombinant vectored vaccines provide economical, rapidly engineered, and tailored immunization strategies. Here, we present a protocol for constructing and characterizing recombinant vectored vaccines for rabies virus (RV). We describe steps for generating replication-competent vesicular stomatitis virus (rcVSV) by replacing vesicular stomatitis virus (VSV) glycoprotein (G) with RV G gene and for recovering rcVSV by co-transfecting baby hamster kidney (BHK-21) cells with the antigenomic VSV cDNA plasmid (pVSV). We then detail procedures for the functional, structural, and molecular characterization of generated rcVSV-RV-Gs.
Source: STAR Protocols
Authors: Manar E. Khalifa, Muhammad Munir
Institution: Lancaster University
Language: English
Subfield: Virology
Field: Immunology and Microbiology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,500
OpenAlex ID: W4403512863