Pseudovirus was prepared
as previously described.46 (link) 293T cells,
seeded 1 day ahead with 2 million cells in 6 cm TC-treated dishes,
were transfected with lentivirus packaging plasmids, SARS-CoV-2 S
plasmid, and lzGreen reporter plasmid.47 (link) After transfection, cells were incubated at 37 °C for 60 h.
Viral media were filtered with a 0.45 μm syringe filter and
snap frozen in liquid nitrogen before storing at −80 °C.
Virus stocks were titrated on 293T-ACE2 cells treated with 50 μL
of 5 μg/mL Polybrene (Sigma-Aldrich, St. Louis, MO, USA). Titer
was determined by fluorescence microscopy using a BZ-X700 all-in-one
fluorescent microscope (Keyence, Itasca, IL, USA).
as previously described.46 (link) 293T cells,
seeded 1 day ahead with 2 million cells in 6 cm TC-treated dishes,
were transfected with lentivirus packaging plasmids, SARS-CoV-2 S
plasmid, and lzGreen reporter plasmid.47 (link) After transfection, cells were incubated at 37 °C for 60 h.
Viral media were filtered with a 0.45 μm syringe filter and
snap frozen in liquid nitrogen before storing at −80 °C.
Virus stocks were titrated on 293T-ACE2 cells treated with 50 μL
of 5 μg/mL Polybrene (Sigma-Aldrich, St. Louis, MO, USA). Titer
was determined by fluorescence microscopy using a BZ-X700 all-in-one
fluorescent microscope (Keyence, Itasca, IL, USA).
