Quantification of rAAV-CMV-Epo vector genomes in the muscles of injected β-thalassemic mice.

(A) The vector diagram: ITR indicates AAV-2 inverted terminal repeats; CMV, immediate early promoter/enhancer elements from CMV; mEpo, murine erythropoietin; and bGH polyA, polyadenylation signal of the bovine growth hormone gene. The BamH-I fragment used as a probe is indicated. (B) Southern blot analysis of BamH-I–digested high-molecular-weight DNA extracted from injected muscle. Analysis was performed after injection at 2 months (mice Nos. 5 and 6), 5 months (mouse No. 11), 10 months (mice Nos. 3 and 10), and 1 year (mice Nos. 1, 2, 4, 7, and 8). Reference copy numbers (ref. copy numbers) correspond to 1 copy (20 pg, lane 1) or 0.1 copy (2 pg, lane 0.1) of the AAV-CMV-Epo plasmid DNA. Endogenous Epo (Epo endo.) and vector signals are shown. Quantification of vector genome copies was performed with a PhosphorImager (Molecular Dynamics) according to the ref. copy number and Epo endo. signals.