Figure 5.

Selective Expansion of Genome-Edited Hepatocytes

Adults or P1 PiZ mice were injected systemically with (A) recombinant AAV8 vector packaged with a promoterless, dual-function cassette containing an artificial miRNA that targets human AAT and a miRNA-detargeted human AAT sequence with a c-Myc tag. The cassette is flanked by 1.1–1.3 kb homology arms to the albumin locus. Transgene expression was evaluated by several methods. (B) Albumin-α-1 antitrypsin-fused transcript/murine albumin transcript ratio by ddPCR. **Statistically significant differences over time (two-way ANOVA). (C) M-AAT-c-Myc serum levels by ELISA, normalized to the average of the age-matched control group. ****Statistically significant differences both over time and between groups (two-way ANOVA). (D) c-Myc immunohistochemistry, quantified by threshold analysis with (E) representative animals shown for each group (scale bar, 2 mm). (D) ****Statistically significant differences over time (two-way ANOVA). For all graphs, data are represented as mean ± SEM.