Figure 5.

Minor transcriptional dysregulation in gene loci targeted by promoter-deprived γ-retroviral self-inactivating (pd-SIN). Real-time reverse transcription–PCR using QuantiTect Primer Assays were used to assess transcription levels of genes adjacent to pd-SIN vector insertions. RNA levels as compared to healthy control mice splenocytes were established using the ΔΔC_t method.^8,49 Only comparatively slight changes in transcription levels were detected. In particular, genes adjacent to insertion sites found to mark clones dominant in more than half of the secondary recipients did not show relevant dysregulation of transcription (I-2: Efcab2; II-3: B3bp). Relative error bars indicate SDs from at least three independent assays.