Homologous recombination mediates stable Fah gene integration and phenotypic correction in tyrosinaemia mouse-model

Figure 1 Flowchart of treating mice. A: Vector map for rAAV8-ROSA26.HAL-TTR.Fah-ROSA26HAR and for rAAV8-TTR.Fah. Fah cDNA is driven by the TTR promotor and for rAAV8-ROSA26.HAL-TTR.Fah-ROSA26HAR located between the homologous arms of the Rosa26 locus. The vector was cloned into an AAV backbone; B: Scheme for the in vivo experiments. First-generation mice (C57BL/6 FAH^∆exon5 strain) were injected with rAAV8-ROSA26.HAL-TTR.Fah-ROSA26HAR (group 1, n = 4) or rAAV8-TTR.Fah (control group, n = 5). The NTBC treatment was stopped, and after 45 d, a partial hepatectomy was performed. In each group, one mouse was used as the donor for hepatocyte transplantation into C57BL/6 Fah^∆exon5 mice. These recipients were the second generation of mice in our study. NTBC treatment was discontinued after hepatocyte transplantation. TTR: Transthyretin promoter (liver specific); R26 HAL: Homologous arm left for target locus in Rosa26; HAR: Homologous arm right for target locus in Rosa26; ITR: Inverted terminal repeat.