Cell and Screening Core

Core B. Cell and Screening Core Summary The focus of the Cell and Screening Core (Core B) is to provide the Projects with cells from diverse mammal species, validated virus stocks, and high throughput screening of virus infections across the "fibroblast zoo" with and without immune perturbation and plasmid and siRNA tools to evaluate virus-host interactions. Viruses need to use host factors to enter cells and complete their replication cycle and need to evaluate antiviral sensing, signaling, and effector protein responses. Given the complexity of these interactions and the potential involvement of many genes and gene networks live cells are needed to fully define mechanisms of species susceptibility and resistance. Fibroblasts are a relevant cell type to achieve these goals because they are target of early infection with alpha and flavivirus and are involved in sensing infection and serve as a source of virus amplification and the site of infection. Fibroblasts are also an ideal cell type for mechanistic studies because they are easy to grow up in large numbers and manipulate in culture and can also be differentiated into other tissue and cell types. This core includes a research team with diverse expertise in virology (Langlois and Cherry), high throughput screening (Cherry), work with primary cells from diverse species (Langlois, Korody, and Maidelaire), and generating stem cells from non-model animals (Korody). Aim 1 will generate primary dermal fibroblasts from diverse mammals for use in the projects and cores. It will also perform validation studies and measuring sources of biological variation. This core will also dedifferentiate fibroblasts from select species into inducible pluripotent stem cells to then differentiate into other cell types for confirmatory studies within the projects. Aim 2 will generate and validate virus stocks for use in the projects and cores. Finally, Aim 3 will perform high throughput screening of alpha and flavivirus infection across the fibroblasts zoo with and without innate immune perturbation or interferon stimulation. Together Core B will provide cells and viruses for the Projects 1 and 2 for mechanistic studies and will provide screening data for Core C and both Projects to further inform mechanisms and to aid in prioritization of virus-host species to evaluate.