Comparison of AAV hFVIII variants. (A) Schematics of 4 rAAV vector genomes encoding hFVIII variants under the control of the LP1 or the smaller HLP regulatory element and upstream of either rabbit globin or synthetic polyadenylation sites. Shown are the A1, A2, A3, C1, and C2 domains of hFVIII either with the SQ B-domain sequences (top schematic) or with the proximal 226-aa region of the B domain (N6). The hFVIII cDNA sequence in the lower 2 constructs are codop. (B) Alkaline gel analysis of viral genomes extracted from 5 × 10¹⁰ rAAV5-LP1-codop-hFVIII-N6 (lane 1) and rAAV5-HLP-codop-hFVIII-N6 (lane 2) viral particles. (C) Mean hFVIII:Ag levels ± SD in murine plasma at 6 weeks after a single tail vein administration of rAAV-hFVIII constructs pseudotyped with serotype 5 capsid in wild-type C57Bl/6 mice (N = 3, dose = 2 × 10¹³ vg/kg). (D) Mean (± SD) proviral copy number in murine liver transduced with rAAV5-hFVIII variants.