Figure 12. Effect of SB100x-mediated integration on the transcriptome of CD34⁺ cells.

(A) Schematic of experiment. CD34⁺ cells were infected with a HDAd5/35++ vector containing a GFP/mgmt cassette under control of the EF1α promoter alone or in combination with HDAd-SB. Transduced cells were expanded in ED medium for 16 days. Two rounds of O⁶BG/BCNU selection (50 μM O⁶BG + 35 μM BCNU) were enriched for GFP-positive cells with integrated transposons. At day 16, GFP-positive cells were FACS sorted (sample 6). For comparison (sample 5), CD34⁺ cells that were transduced with the mgmt/GFP vector alone and subjected to selection were used. Because the control cells did not express SB100x, they lost the episomal mgmt/GFP vector and were therefore GFP negative. Total RNA from both samples was subjected to RNA-Seq performed by Omega Bioservices. (B) Genes with altered mRNA expression (log₂ fold change) ranked based on their P value.