Figure 4.

Analysis of gene transfer efficiency by quantitative PCR in liver, spleen, and lungs (A) and in testis, heart, brain, kidney, stomach, and intestine (B). The average vector copy number per diploid genomic equivalent was determined by comparison with a serially diluted linear standard (correlation coefficient r² = 0.98) ranging from 0.5 to 0 (negative control) copies per diploid genomic equivalent. Organs from individual mice were indicated (1, 2, 5, and 7 from Fig. 1 and Table 1). Mice 1 and 2 were not killed, but a liver biopsy was taken instead, and decreasing amounts of target DNA were used as template: 200 ng (a), 100 ng (b), and 50 ng (c). A constant amount of total DNA (200 ng) was maintained in the standard and in the liver samples from mice 1 and 2 by adding spleen DNA from a FVIII-deficient mouse. FVIII (1.1 kb) or control β-actin (0.2 kb)-specific fragments are indicated. The molecular weight (MW) marker corresponds to the Smart Ladder.