Inadvertent Transfer of Murine VL30 Retrotransposons to CAR-T Cells

Figure 5

The effect of 293T cell replication on GCN of HIV-1 vector-transferred VL30. (a) Bar graph showing VL30 GCN in 293T cells treated with conditioned media containing HIV-1 vector particles packaged with VL30 genomes. Conditioned media samples were collected from cell clones 6 and 7 following transient transfection with a second-generation HIV-1 vector packaging plasmid and a VSV-G envelope expression cassette. DNA samples of target 293T cells were analyzed by qPCR for the presence of VL30 genomes either at 24 h posttransduction (P0) or following 4 passages (P4). values were determined by the 2-way ANOVA test. The experiment was performed in triplicate. (b) Electrophoresis analysis of PCR products following amplification of DNA samples extracted from 293T cells transduced by HIV-1 particles generated in cell clones 6 and 7. DNA samples extracted from 293T cells transduced with lentiviral vectors pTK1261 and pTK2229 served as negative and positive controls, respectively, and are shown as –Con and +Con, respectively. PCR amplification products of the endogenous RNaseP gene served as loading controls.

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