ATeam technology for detecting early signs of viral cytopathic effect

Abstract

24Adenosine 5'-triphosphate (ATP), the major energy currency of the cell, is involved in 25 many cellular processes, including the viral life cycle, and can be used as an indicator of early 26 signs of cytopathic effect (CPE). In this study, we demonstrated that CPE can be analyzed using 27 an FRET-based ATP probe named ATP indicator based on Epsilon subunit for Analytical 28Measurements (ATeam). The results revealed that as early as 3 hr, the virus infected cells showed 29 a significantly different Venus/cyan fluorescent protein (CFP) ratio compared to the mock-infected 30 cells. The ATeam technology is therefore useful to determine the early signs of ATP-based CPE 31 as early as 3 hr without morphology-based CPE by light microscopy, and enables high throughput 32determination of the presence of microorganisms in neglected samples stored in laboratories. 33 34

An expanded palette of ATP sensors for subcellular and multiplexed imaging

“…Genetically encoded fluorescent protein-based biosensors of ATP have been essential to illuminating the real-time dynamics of ATP regulation in live cells. Biosensors such as ATeam1.03 and iATPsnFR have been used to elucidate cell-type specific metabolic differences and interactions [39][40][41][42] , the spatiotemporal correlation between ATP and other signaling molecules like calcium, sodium, and insulin [43][44][45][46] , and previously undefined roles of ATP in processes such as viral replication and cytopathic effect 47,48 . These sensors have also been used to image changes in extracellular ATP concentrations, enabling characterization of ATP as a signal mediating cell-to-cell communication 13,49,50 .…”