Fig. 2: Designed AAV_ITGs were well-produced and improved transduction via αVβ6 binding.
From: An engineered AAV targeting integrin alpha V beta 6 presents improved myotropism across species
A AAV titers of different AAV variants in bulked small-scale production in suspension 3-day post-triple-transfection (2 ml production, 6 biological replicates, one-way ANOVA followed by FDR correction). B Western blot of VP proteins from purified AAVs showed similar VP ratios for designed AAV_ITGs capsids compared to AAV9 and AAV9rh74, suggesting successful capsid assembly. C, D VCN (C) and luciferase activity (D) of 293_αVβ6 after AAV infection (3 and 4 biological replicates, one-way ANOVA followed by FDR correction). Both the two designed AAV_ITGs showed enhanced VCN and luciferase activities compared to AAV9rh74 and AAV9. E Inhibition of cell entry of designed AAV_ITGs, but not for AAV9 or AAV9rh74, in 293_αVβ6 cells by αVβ6 recombinant protein. AAVs were preincubated with human αVβ6 recombinant protein (r.ITGAV-B6) for 30 min at 37 °C before infection (4 biological replicates, two-way ANOVA followed by FDR correction, 1 µg protein per 5E9vg AAV, 2E5 vg per cell). The same condition treated with recombinant SGCA protein (r.SGCA) was used as the control. F–J Enhanced transduction of AAV_ITGs in in vitro human differentiated myotubes, but not in myoblasts. F Representative images of the GFP signal of myotubes 48 h post-infection (scale bar: 400 µm). G–J VCN and luciferase activities of AAV_ITGs in comparison with AAV9 and AAV9rh74 in myoblasts (G, I, 3 and 3/4 biological replicates, respectively) and myotubes (H, J, 3 and 3/4/6 biological replicates, respectively) (one-way ANOVA followed by FDR correction). Data are presented as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns not significant. Source data are provided as a Source Data file.
