Biodistribution of scAAV2/5-LP1-hFIXco following peripheral vein administration of scAAV2/5. (A) One microgram of genomic DNA, isolated from the indicated organs 4 weeks after peripheral vein administration of 1 × 10¹² vg/kg scAAV2/5 particles in M8-sc (top set of panels) and M9-sc (bottom set of panels), was subjected to PCR using primers unique to hFIXco designed to amplify a 617 bp product. Integrity of the DNA was determined by amplifying a 604 bp region of the rhesus β-actin gene and is shown at the bottom of these panels. The middle panel is RT-PCR analysis of RNA extracted from the organs of M8-sc. Integrity of the RNA was determined by amplifying a 295 bp region of the rhesus GAPDH gene and is shown at the bottom of the panel. (B) Q-PCR reactions were performed in duplicate on genomic DNA to establish transgene copy number in each organ after peripheral (⊡, n = 3) and mesenteric (□, n = 2) vein administration of scAAV2/5. The results are represented as vector copy number per diploid genome together with standard errors of mean.