Lentivirus and MLV vectors transduce HSCs with similar efficiency.

Representative co-transduction assay of Lin⁻ mouse BM cells co-transduced with MLV-GFP-VSV-G (M-GFP) and ecotropic-MLV-LacZ (MLV group) or lentivirus-GFP-VSV-G (L-GFP) and ecotropic-MLV-LacZ (lenti group). Lin⁻ mouse BM cells were transduced for 24 hours without cytokines with medium changed at 0 and 12 hours or for 96 hours in IL-3, IL-6, and SCF with medium changed every 24 hours. After transduction, the cells were injected intoW/W^v recipients. Peripheral blood DNA from the recipients was analyzed 16 weeks after transplant for the presence of the proviruses by PCR, using GFP- and LacZ-specific primers. PCR of the VSV-G-EGFP provirus generates a 586-bp product, and PCR of the ecotropic-lacZ provirus generates a 289-bp product. DNA from the producer cell lines was used as a positive control, and DNA from an untransplanted mouse was used as a negative control. The mouse adult β-globin gene was amplified as an internal control. Phosphor Imager analysis confirmed that each sample had equivalent amounts of DNA.