Project description:To better prepare for future viral outbreaks, scalable and adaptable platforms to study emerging infections are essential. Understanding virus–host interactions, particularly the mechanisms of cell entry, is critical for developing effective therapeutics and vaccines. Current approaches often rely on live virus assays requiring high-containment facilities, limiting speed, scalability, and accessibility. As a proof-of-principle, we developed a novel screening platform—Ceudovitox—using pseudotyped viruses (PVs) bearing the chikungunya virus (CHIKV) envelope protein. These PVs were engineered to express herpes simplex virus-1 thymidine kinase, enabling selective killing of infected cells with ganciclovir. A heterogeneous CRISPR-Cas9 knockout cell pool was then screened using this "killer" PV system, allowing identification of CHIKV entry factors via next-generation sequencing.
Project description:The purpose of this experiment was to obtain samples for mRNA analysis in IHH cells infected with Zaire Ebola virus and mutants: Zaire Ebola virus: This wild-type Ebola virus - strain Mayinga - was isolated from a fatal human case in Zaire (now known as the Democratic Republic of Congo) in 1976 Zaire Ebola virus, VP35 R312A possesses a R312A mutation in the VP35 protein. Zaire Ebola virus, delta sGP. Lacks the ability to produce non-structural protein, the secreted glycoprotein (sGP). Zaire Ebola virus, delta mucin. Lacks the mucin-like domain (MLD), which contains both N-linked and O-linked glycosylation sites, for the glycoproteins.
Project description:Dengue and Zika are closely related members of the Flaviviridae family of positive, single-stranded RNA viruses and are of global clinical importance. These viruses utilize an 11kb RNA genome for translation and replication, and much remains to be learnt about how the entire genome folds to enable virus function. Here, we performed high throughput RNA secondary structure and pair-wise interaction mapping on four dengue serotypes and four Zika strains within their virus particles. We identified structures that are associated with translation pausing, and are evolutionary conserved by integrating synonymous mutation rates into our analysis. Genome-wide interaction mapping revealed alternative structures, as well as extensive long-range RNA interactions – including the known circularization signals– within the virus particles. Many of these long-range interactions are conserved across the viruses and/or clustered into “hubs” that are shown to be functionally important. This comprehensive structural resource of dengue and Zika viruses reveals that viral genome organization is much more complex than previously appreciated and deepens our understanding of the molecular basis for viral pathogenesis.