Fig. 1

CHSE salmon cells are efficiently transduced with lentivirus. a Efficient transduction of CMV:EGFP in CHSE Chinook salmon cell line by lentivirus. Salmon cells were spinfected at 22 °C with neat lentivirus supernatant for 2 h at 1000 x g. The cells were incubated for 24 h and the media was replaced. After 2 weeks of expansions, fluorescence was recorded by flow cytometry using CHSE wt (not transduced) as control. Split histogram of control cells (top) and pLenti-GFP transduced CHSE with optimal conditions (bottom). Data were normalised to Mode (relative percentage of cells rather than number). b Representative image of CHSE cells 8 days post transduction with pLenti-GFP. Differential Interference Contrast (top) and GFP (bottom) channels are represented. Arrows point to GFP negative cells. Scale bar 20 μm. c-d Different conditions of the optimisation protocol with optimal conditions in purple. c, Incubation temperature and spinfection influences the efficiency of transduction. Cells were transduced with neat lentivirus supernatant, spinfected for 2 h at 1000 x g (or not) and incubated at 17 °C (17), 22 °C (22) or heat shocked for 4 h at 28 °C followed by 22 °C for 2 weeks (28–22) until flow cytometry. d, Incubation of the cells with the virus for 4 h reduces the efficiency of transduction. CHSE were transduced with neat lentivirus dose at different temperatures and media was changed after 4 or 24 h