Approaches for genetic engineering ofrhabdoviruses such as VSV and paramyxoviruses such as SV5 haveproven to be more complex than those used for many other RNA viruses. This is due to inherent properties ofthe negative strand RNA virus genome and growth cycle, both of which present challenges to construction ofdefined vectors that are not solved by traditional recombinant DNA approaches. However, recent state-of-the-art technologies have been developed to overcome these obstacles and these approaches now allow thegeneration of recombinant negative strand RNA viruses containing pre-determined alterations to theirgenomes. The Virology Core Laboratory of the Program will function as a resource to carry out these newapproaches in order to facilitate the generation and purification of novel recombinant viruses for Projects 2and 3. In addition, the Core will function as a source of purified virus preparations that are of a consistenthigh quality for use in the vaccination experiments propose in all four Projects.The goal of the Virology Core Laboratory of the Program is to perform a consistent, uniform and highquality set of tasks needed by each of the four Projects, including recovery of recombinant viruses,defining growth and gene expression properties, and providing viral stocks for vaccination of mice.
In aim1, the Core will use state-of-the-art technologies to facilitate the rapid and consistent recovery ofrSV5 andrVSV vectors from engineered plasmid DNA.
In aim 2, the Core will carry out assays for single step andmulti-step growth, establishing viral RNA synthesis profiles and determining the properties of foreign geneexpression.
In aim 3, the Core will carry out the purification of virus stocks, titrating of stocks and qualitycontrol assays for the levels of contaminationCompletion of these tasks will assure a reproducible and dependable source of viral vectors for the ProgramProject such that comparisons can be made across Projects.
Showing the most recent 10 out of 25 publications
