Ebola virus is an emerging human pathogen that is associated with a severe hemorrhagic fever and remains a threat due to its possible use as a bioterrorism agent. Currently, there are neither vaccines, nor antivirals available to prevent or treat Ebola virus infections. Our laboratory has successfully used virus like particles (VLPs) to study the role of Ebola virus matrix protein VP40 late domains in VP40 VLP budding. We have also described a role for other viral and host components in VP40 VLP budding. Based on our considerable experience investigating VP40 VLP budding we hypothesize that our VP40 VLP budding assay can be utilized to identify the packaging signal of the Ebola virus 3E-5E minigenome RNA, and thus could serve as a platform for a VLP based vector for vaccine development or as a gene therapy vector. Our preliminary data indicate that VP40 and VP35 are the minimal viral proteins required to package the 3E-5E minigenome RNA into budding VLPs. We propose to identify the Ebola virus RNA packaging signal by generating mutants of the 3E-5E minigenome and test them in our modified VP40 VLP budding assay. We further aim to identify regions of VP35 required for packaging and interactions with the 3E-5E minigenome RNA as well as interactions with VP40. Once the packaging signal is identified, a model RNA will be generated containing the packaging signal as well as a protein of interest such as red fluorescent protein. We will then test our VLP for its ability to transduce cells in tissue culture and produce protein from our model RNA. Development of such a system would increase our understanding of the molecular requirements of Ebola virus genome packaging and could be used as an antigen delivery platform to produce an efficacious humoral and cellular immune response. ? ? ?
| Harty, Ronald N (2009) No exit: targeting the budding process to inhibit filovirus replication. Antiviral Res 81:189-97 |
