cag promoter pscaav cag gfp  (Addgene inc)

Journal: Liver Research

Article Title: Efficient AAV8 delivery to the liver via isolated hepatic perfusion and analysis of hepatic lobule transduction patterns

doi: 10.1016/j.livres.2025.04.006

Figure Lengend Snippet: IHP efficiently delivers AAV8 to each lobe of the rat liver. (A) Fluorescence distribution in each liver lobe. (B) Area fraction of dTomato + cells in each liver lobe. (C) Average fluorescence intensity of dTomato + cells in each liver lobe. L, left lateral; LM, left medial; M, middle; RL, right lower; RU, right upper; T, tail. (D) Representative images of hepatic cell marker FAH and dTomato co-staining in areas with dTomato high and low expression levels. Five images per rat ( n = 4) used for statistical analysis. ∗∗∗∗ P < 0.0001. Abbreviations: AAV8, adeno-associated virus 8; DAPI, 4′,6-diamidino-2-phenylindole; FAH, fumarylacetoacetate hydrolase; IHP, isolated hepatic perfusion.

Article Snippet: All AAV8 vectors expressing dTomato protein under a CAG promoter were purchased from PackGene Biotech (Guangzhou, China), with an initial titer of 10 13 genome copies (GC)/mL.

Techniques: Fluorescence, Marker, Staining, Expressing, Virus, Isolation

Journal: Liver Research

Article Title: Efficient AAV8 delivery to the liver via isolated hepatic perfusion and analysis of hepatic lobule transduction patterns

doi: 10.1016/j.livres.2025.04.006

Figure Lengend Snippet: AAV8 delivery by IHP exhibits partial dose dependence and uneven distribution characteristics in the rat liver. (A) Fluorescence distribution under different dose settings (2 × 10 12 GC/kg, 4 × 10 12 GC/kg, and 7 × 10 12 GC/kg, respectively). (B) Area fraction of fluorescence expression in the unit field of view of liver tissue slices under different dose settings. (C) Mean fluorescence intensity in the unit field of view of liver tissue slices under different dose settings. (D) Representative images of hepatic cell marker FAH and dTomato co-staining under different dose settings. (E) The proportion of FAH and dTomato co-stained cells in FAH + hepatocytes under different dose settings. n = 4 per group, ∗ P < 0.05, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. ns, not significant. Abbreviations: AAV8, adeno-associated virus 8; DAPI, 4′,6-diamidino-2-phenylindole; FAH, fumarylacetoacetate hydrolase; GC, genome copies; IHP, isolated hepatic perfusion.

Article Snippet: All AAV8 vectors expressing dTomato protein under a CAG promoter were purchased from PackGene Biotech (Guangzhou, China), with an initial titer of 10 13 genome copies (GC)/mL.

Techniques: Fluorescence, Expressing, Marker, Staining, Virus, Isolation

Journal: Liver Research

Article Title: Efficient AAV8 delivery to the liver via isolated hepatic perfusion and analysis of hepatic lobule transduction patterns

doi: 10.1016/j.livres.2025.04.006

Figure Lengend Snippet: IHP can achieve higher intrahepatic AAV8 delivery efficiency than PV and IVC in the rat liver. (A) Fluorescence distribution under different delivery AAV8 vector routes. (B) Area fraction of dTomato fluorescent expression under different delivery routes. (C) Mean fluorescence intensity under different delivery routes. n = 4 per group, ∗∗∗ P < 0.001. ns, not significant. Abbreviations: AAV8, adeno-associated virus 8; DAPI, 4′,6-diamidino-2-phenylindole; IHP, isolated hepatic perfusion; IVC, inferior vena cava; PV, portal vein.

Article Snippet: All AAV8 vectors expressing dTomato protein under a CAG promoter were purchased from PackGene Biotech (Guangzhou, China), with an initial titer of 10 13 genome copies (GC)/mL.

Techniques: Fluorescence, Plasmid Preparation, Expressing, Virus, Isolation

Journal: Liver Research

Article Title: Efficient AAV8 delivery to the liver via isolated hepatic perfusion and analysis of hepatic lobule transduction patterns

doi: 10.1016/j.livres.2025.04.006

Figure Lengend Snippet: The IHP route makes AAV8 vectors present a highly concentrated distribution near the zone 1 area of the rat liver. (A) Representative image of uneven distribution of dTomato + cells. The gray-bordered squares represent areas of high expression of dTomato, while white borders represent areas with relatively low expression of dTomato. (B) Magnified view of a field with high expression of dTomato. (C) Magnified view of a field with low expression of dTomato. (D, E) Representative images show the colocalization of dTomato and GS immunofluorescence staining at different magnifications (D: low-power images; E: high-power images). (F) Schematic diagram of transduction efficiency analysis around portal and central veins. The hexagons represent the liver lobules, and the red and black circles represent the portal and central veins in the center of the field of view. Circles' diameter is 450 μm. (G) No statistical difference was observed in the area fraction of portal/central transduction efficiency among different delivery routes. (H) IHP delivery showed a statistically significant difference in fluorescence intensity for portal/central transduction efficiency compared to the other two routes, indicating a higher infection intensity near the portal vein. n = 4 per group, ∗ P < 0.05, ∗∗ P < 0.01. ns, not significant. Abbreviations: AAV8, adeno-associated virus 8; DAPI, 4′,6-diamidino-2-phenylindole; GS, glutamine synthetase; IHP, isolated hepatic perfusion; IVC, inferior vena cava; PV, portal vein.

Article Snippet: All AAV8 vectors expressing dTomato protein under a CAG promoter were purchased from PackGene Biotech (Guangzhou, China), with an initial titer of 10 13 genome copies (GC)/mL.

Techniques: Expressing, Immunofluorescence, Staining, Transduction, Fluorescence, Infection, Virus, Isolation

Journal: Liver Research

Article Title: Efficient AAV8 delivery to the liver via isolated hepatic perfusion and analysis of hepatic lobule transduction patterns

doi: 10.1016/j.livres.2025.04.006

Figure Lengend Snippet: Delivery of AAV8 to cynomolgus monkey liver through IHP route. (A) The surgical images before and after perfusion are presented. The two images on the left side show black arrows indicating the blood vessels to be clamped, including the inferior vena cava and the portal vein. (B) Statistical diagram of AAV8 transduction efficiency in different liver lobes. (C) Representative images illustrating the distribution of AAV8 in different liver lobes. L, left lateral; LM, left medial; M, middle; R, right; T, tail. (D, E) Representative images show the colocalization of dTomato and GS immunofluorescence staining at different magnifications (D: low-magnification images; E: high-magnification images). (F) Representative images show the distribution of dTomato + cells in the heart, brain, spleen, lung, and kidney. (G) Representative histological images of liver tissue at post-IHP surgery. (H) Representative histological images of right kidney tissue at post-IHP surgery. ( n = 1). ∗∗∗ P < 0.001. Abbreviations: AAV8, adeno-associated virus 8; DAPI, 4′,6-diamidino-2-phenylindole; GS, glutamine synthetase; IHP, isolated hepatic perfusion.

Article Snippet: All AAV8 vectors expressing dTomato protein under a CAG promoter were purchased from PackGene Biotech (Guangzhou, China), with an initial titer of 10 13 genome copies (GC)/mL.

Techniques: Transduction, Immunofluorescence, Staining, Virus, Isolation