HEK293T embryonic kidney cells (CRL‐3216), and 143B osteosarcoma cells (CRL‐8303) were obtained from ATCC. HEK293 Flp-In T-REx 293 cells were obtained from Invitrogen. The 143B.TK− rho0 derivative (143B206), obtained from Dr Carlos Moraes, was generated by Dr M.P. King (24 (link)). The details of the cell lines are listed in Supplementary Table S4 . HEK293T cells KO for MTG1, GTPBP10, or DDX28 were previously generated and reported by our group (4 ,9 (link),11 (link)). All cell lines were cultured at 37°C under 5% CO2 in high-glucose, sodium pyruvate, l -glutamine DMEM medium supplemented with 10% FBS and 100 μg/ml uridine (complete DMEM medium). Analysis of mycoplasma contamination was routinely performed. The chemicals and reagents used in this study are listed in the Supplementary Table S8 .
Protocol detail
Characterization of Engineered Cell Lines
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Cell lines
Cells
Embryonic
Glucose
Kidney
L glutamine
Mycoplasma
Osteosarcoma
Pyruvate
Sodium
T cells
Uridine
Corresponding Organization : University of Miami
Other organizations : McGill University, Montreal Neurological Institute and Hospital, Mount Sinai Hospital, Lunenfeld-Tanenbaum Research Institute, University of Toronto
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Variable analysis
independent variables
- Knockout of MTG1, GTPBP10, or DDX28 genes in HEK293T cells
- Not explicitly mentioned
- Cell culture conditions: 37°C, 5% CO2, high-glucose, sodium pyruvate, L-glutamine DMEM medium supplemented with 10% FBS and 100 μg/ml uridine
- Mycoplasma contamination analysis routinely performed
- Positive control: HEK293T, 143B osteosarcoma, and HEK293 Flp-In T-REx 293 cell lines
- Negative control: 143B.TK- rho0 derivative (143B206) cell line
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