Oncolytic Reovirus mediates innate-driven SARS-CoV-2 elimination in the absence of cell toxicity
Oncolytic Reovirus mediates innate-driven SARS-CoV-2 elimination in the absence of cell toxicity
Cells were washed three times in PBS and lysed with TRIzol according to manufacturers instructions. cells were cultured at 37 Â°C in a humidified atmosphere at 5 % CO2 in Dulbecco's modified Eagle medium (DMEM) containing high glucose and supplemented with 10 % (v/v) heat-inactivated foetal bovine serum (FBS) and 1 % (v/v) non-essential amino acids. A549 AT cells were infected with SARS-CoV-2 (Eng/2) at 0.5 PFU/cell. Reo were administered either 12 h before SARS-CoV-2 infection (pretreatment), at 0 hpi (same time), or 6 hpi (5 PFU/cell). Cells were incubated for 24 h following SARS-CoV-2 infection. RNA was purified using TRIzol according to manufacturers instructions. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using either dUTP for directional library or dTTP for non-directional library. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. 