Nucleic acid-based vaccine delivery platforms have recently proven highly successful for effective control of infectious diseases. This was highlighted by the extreme success of lipid nanoparticle (LNP)-based mRNA vaccines for SARS-CoV-2 which curbed the number of hospitalizations due to COVID-19 and led to control of the COVID-19 pandemic. Despite the success in human health mRNA-based platforms present several drawbacks that hinder their immediate use and broad application for vaccine delivery in veterinary medicine, including the cost of production, the low stability of mRNA and the need for ultralow temperature storage. DNA-based vaccine delivery platforms such as linearDNA overcome some of these limitations and may offer the ideal nucleic acid-based platform for vaccine delivery in animal species. We know focus on delivery methods for linear DNA beyond electroporation. In the proposed project, we will address this limitation and optimize LNP-based formulations for delivery of linearDNA vaccine candidates. The objectives of the proposed project are:

Objective 1 (O1): To optimize formulations and screen LNP-formulations for efficient delivery of linearDNA-based platforms encoding reporter genes (i.e. GFP and luciferase) in cell lines from multiple animal species in vitro and in a mouse model in vivo.

Objective 2 (O2): To design and optimize expression and nuclear delivery of candidate antigens/payloads of viral pathogens affecting horses (i.e. equine influenza virus [EIV]).

Objective 3: To assess immunogenicity of linearDNA-LNP platform encoding the selected equine viral antigens in a mouse model in vivo.