Activity of rAAV-hFIX vectors in murine livers.
In vivo activity of rAAV CAGG-FIX (n = 6; A) was compared with the 3 other vector constructs (B: CMV, solid squares, n = 5; MSCV, open squares, n = 5; HBP, triangles, n = 6) following a single injection of 1 × 1011 vector particles into the portal vein of 7- to 10-week-old male C57Bl/6 SCID mice. Mice were bled at the indicated number of weeks following vector administration and assayed for hFIX, using an ELISA. The results are represented as means with standard error. (C) Southern blot analysis of total DNA isolated from the liver of mice 22 weeks following administration of 1 × 1011vectors particles of each rAAV-hFIX vectors. DNA samples were digested with EcoRI, transferred to nitrocellulose membrane by Southern blot procedure, and then hybridized with a 32P radiolabeled 1.5-kb hFIX cDNA probe. From right to left, lane 1 = markers, lane 2 = rAAV-FIX construct containing the MSCV promoter, lane 3 = CAGG promoter, lane 4 = CMV promoter, lane 5 = HBP promoter, lane 6 = DNA from a mock transduced mouse, and lanes 7 to 9 = control mouse DNA spiked with 10, 1, and 0.1 copies per diploid genome of plasmid pAV CAGG-FIX digested withEcoRI.
