Fig. 1: Generation of the TIRFA strain.
From: A humanized mouse model for adeno-associated viral gene therapy
a AAVR gene was knocked out in TIRF zygotes using CRISPR/Cas9 gene editing tools. b AAVR gene scheme showing the sgRNAs used to target exons 4 and exons 5 and the primers binding sites used to screen the mice. On the right, DNA gel electrophoresis shows PCR bands of heterozygous and homozygous mice. c, d Human albumin levels of humanized TIRF control and TIRFA mice transplanted with two different hepatocyte donors (n = 18 for TIRF donor#1, n = 23 for TIRF donor#2, n = 7 for TIRFA donor#1 and n = 14 for TIRFA donor#2) c and its increase over time d (n = 3 for TIRF donor#1, n = 4 for TIRF donor#2, n = 3 for TIRFA donor#1 and n = 6 for TIRFA donor#2), e FAH immunostaining and H&E of TIRF and TIRFA humanized mice livers f tdTomato immunostaining of different organs from a TIRFA mouse injected with AAV8-tdTomato virus. On the right, amplification of the heart tissue. g Immunofluorescence staining of the livers from TIRF and TIRFA humanized mice injected with AAV8 virus expressing tdTomato (red). Human cells are stained with hLDH (green). Results observed in (b) and (e–g) were observed in at least 3 independent samples per condition. sgRNA single guide RNA, PCR polymerase chain reaction, H&E Hematoxylin and Eosin, FAH fumarylacetoacetate hydrolase, AAV adeno-associated virus, RFP red fluorescent protein, hLDH human lactate dehydrogenase. Data are presented as means ± SD. Significance was validated with a two-sided student t-test. Source data are provided as a Source Data file. Scale bars represent 50, 100, or 200 μm, respectively.
