Additionally, by combining different HCV genotypes, enables to identify drug candidates with cross-genotypic coverage and allowstriaging of potentially
genotype-specific compounds. Finally, the advantage of monitoring cytotoxic effects in parallel reduces the probability of selecting less favorable compounds. Selleckchem CX-5461 Taken together, the phenotypic assay described here facilitates the selection of antivirals with a novel mechanisms of action, which are potential new therapeutics and tools to elucidate the still poorly understood HCV life cycle. This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST/No. 2011-00244), Gyeonggi-do and KISTI. Selleck AT13387 C.T.J. and C.M.R. were supported by grants from the NIH (CA057973 and DK085713), the Starr Foundation and the Greenberg Medical Research Institute. “
“Ebolaviruses are non-segmented negative sense RNA viruses in the family Filoviridae. Ebola virus (EBOV) causes a severe hemorrhagic fever in humans and non-human primates with case fatality rates in humans of up to 90% ( Feldmann and Geisbert, 2011). Despite intensive research, there are no approved therapies available for treatment of Ebola hemorrhagic fever ( Kondratowicz and Maury, 2012). One factor that has hindered the development of efficient therapies is the fact that wild-type
EBOV is not very amenable to antiviral screening, which is at least in part due to the fact that development of cytopathic effect (CPE), Loperamide which is the easiest way to detect infection, is relatively slow ( Pegoraro et al., 2012). Reverse genetics systems allow the generation of recombinant EBOVs (Hoenen et al., 2011), and have been used in the past to generate eGFP-expressing
EBOVs (Ebihara et al., 2007 and Towner et al., 2005), which allow much more rapid detection of infection in vitro. Using these viruses great progress has recently been made in developing high-content screening protocols for EBOV ( Panchal et al., 2010 and Pegoraro et al., 2012). However, high-content screening requires extensive and costly automated imaging equipment, and so far these protocols have relied on a multistep approach in which cells are first infected in a BSL4 laboratory for several days, and then fixed for several days in formalin before they are analyzed under BSL2 conditions ( Panchal et al., 2012 and Pegoraro et al., 2012). Luminescent reporters provide a viable alternative to fluorescent reporters (Miraglia et al., 2011). They facilitate very sensitive cell-based reporter assays (Thorne et al., 2010), eliminate the problem of compound fluorescence (Simeonov et al., 2008), and have relatively modest instrumentation requirements. Therefore, as an alternative to the eGFP-expressing EBOV, we have developed a recombinant EBOV expressing Firefly luciferase (rgEBOV-luc2) as a reporter protein.