biostudies-literatureRobinson SMNIAID NIH HHSNHLBI NIH HHSNIMH NIH HHSNINDS NIH HHSe1004045https://www.ebi.ac.uk/biostudies/studies/S-EPMC3983045biostudies-literatureUnknown10(4)Coxsackievirus B3 (CVB3), a member of the picornavirus family and enterovirus genus, causes viral myocarditis, aseptic meningitis, and pancreatitis in humans. We genetically engineered a unique molecular marker, "fluorescent timer" protein, within our infectious CVB3 clone and isolated a high-titer recombinant viral stock (Timer-CVB3) following transfection in HeLa cells. "Fluorescent timer" protein undergoes slow conversion of fluorescence from green to red over time, and Timer-CVB3 can be utilized to track virus infection and dissemination in real time. Upon infection with Timer-CVB3, HeLa cells, neural progenitor and stem cells (NPSCs), and C2C12 myoblast cells slowly changed fluorescence from green to red over 72 hours as determined by fluorescence microscopy or flow cytometric analysiPLoS pathogensCoxsackievirus B exits the host cell in shed microvesicles displaying autophagosomal markers.PMC3983045R24 MH065515F32 AI065095R01 AI0423143R01NS054108-01A2S1R01 HL092136R01 NS054108F32 AI-065095T32 HL116273R01 HL093177Sin JDoran KSCornell CTMangale VKha NCruz CSayen MRHancock BMRahawi SNguyen DPRobinson SMSegall AMTsueng GGottlieb RAMcIntyre LLWilliams WWhitton JLWolkowicz RFeuer RHilton BJfalseCoxsackievirus B exits the host cell in shed microvesicles displaying autophagosomal markers.Coxsackievirus B3 (CVB3), a member of the picornavirus family and enterovirus genus, causes viral myocarditis, aseptic meningitis, and pancreatitis in humans. We genetically engineered a unique molecular marker, "fluorescent timer" protein, within our infectious CVB3 clone and isolated a high-titer recombinant viral stock (Timer-CVB3) following transfection in HeLa cells. "Fluorescent timer" protein undergoes slow conversion of fluorescence from green to red over time, and Timer-CVB3 can be utilized to track virus infection and dissemination in real time. Upon infection with Timer-CVB3, HeLa cells, neural progenitor and stem cells (NPSCs), and C2C12 myoblast cells slowly changed fluorescence from green to red over 72 hours as determined by fluorescence microscopy or flow cytometric analysi2014-01-01T00:00:00Z2014 Apr2025-04-18T17:36:37.753Z2019-03-27T01:24:47ZS-EPMC39830452472277310.1371/journal.ppat.1004045