FIG. 7.

Analysis of AAV2 lysine mutant vector-mediated EGFP expression in hepatocytes of normal C57BL/6 mice in comparison with wild-type AAV2 vector-mediated EGFP expression. (A) Transgene expression was detected by fluorescence microscopy 4 weeks post-injection of scAAV2-EGFP or AAV2 K→R mutant vector at 5×10¹⁰ vector particles per animal. Representative images of hepatic tissues from four different animals in each group are shown. (B) Estimation of vector genome copies in liver after AAV-mediated gene transfer. Genomic DNA was isolated from the liver tissue of C57BL/6 mice 4 weeks after vector administration and the viral copy numbers were estimated by quantitative PCR as described in Materials and Methods. (C) Analysis of EGFP transcript levels by real-time quantitative PCR. Hepatic RNA isolated from animals injected with AAV2-WT or K→R mutant vector was analyzed for EGFP expression; the data are normalized to the GAPDH reference gene. One-way analysis of variance (ANOVA) was used for the statistical comparisons. *p<0.05 versus AAV2-WT-injected animals. Color images available online at www.liebertpub.com/hgtb