Figure 3

The effect of rAAV2-TTR-RB15-mediated gene expression in LDLR-/-Apobec1-/- mice. (A). The DNAs from various organs were extracted at days 7 and 150 after rAAV2-TTR-RB15 transduction. The AAV2-RB15 DNA was detected by amplification using PCR, followed by Southern blot analysis. The radioactive bands of RB15 are shown. (B). The expressed ribozymes RB15 and RB15 mutant RNAs in the livers of LDLR-/-Apobec1-/- mice treated with rAAV2-TTR-RB15 or RB15-mutant. The total liver RNA was extracted from each mouse at day-150 after rAAV-TTR-RB15 (2 × 10¹¹ particles/animal, samples 1 – 5) or rAAV-TTR-RB15 mutant (2 × 10¹¹ particles/animal, samples 6 – 10) transduction. Ribozymes RB15 or RB15 mutant RNAs were detected using RT-PCR, followed by Southern blot analysis. The radioactive bands of ribozymes RB15 and RB15 mutant are shown. The same samples were also subjected to PCR only, followed by Southern blot analysis. There were no detectable bands in these samples. (C). The effect of ribozymes RB15 and RB15 treatment on mouse apoB mRNA levels. Mouse apoB mRNA and endogenous 18S RNA from each mouse liver at day-150 after rAAV-TTR-RB15 or RB15 mutant treatment were determined by real-time quantitative RT-PCR. The results are expressed as the ratio of mouse apoB mRNA (mApoB mRNA) to 18S RNA. The results are shown as means ± standard deviation of ten animals. Each assay was performed in duplicate at two separate times. The results were analyzed by Student t-test.