High-level β-globin expression and preferred intragenic integration after lentiviral transduction of human cord blood stem cells
J. Clin. Invest. Suzan Imren, et al. 114:953 doi:10.1172/JCI21838 [Go to this article.]

Figure 4
Bubble LM-PCR analysis of the number and dynamics of β^A-T87Q -globin lentivirus-positive clones in mice transplanted with transduced cord blood cells. (A_C) Analysis of individual CFC-derived colonies from 3 recipients (A, 1 recipient from experiment 4 analyzed 24 weeks after transplant; B and C, 2 mice from experiment 2, described in Figure 2A, analyzed 19 weeks after transplant). DNA from a transgene-negative colony was used as a negative control for the PCR, and DNA from a mouse spleen colony with a single integration confirmed by Southern blotting was used as a positive control. Note that the expected 750-bp 3′ LTR_related fragment (IB) could not be detected in all of the colonies. The presence of identical faint bands in the analyses of several colonies is probably caused by cross contamination that occurred while plucking these colonies from the methylcellulose cultures. (D and E) Analysis of CFC-derived colonies from a third mouse from experiment 2 described in Figure 2A analyzed 11 and 19 weeks after transplant. Bands with asterisks were gel purified and sequenced. Asterisks with the same color indicate colonies that showed the same integration site upon sequencing.