AAV plasmids containing different versions of the G protein gene were submitted to the Viral Vector Core of The Research Institute at Nationwide Children’s Hospital (Columbus, Ohio, USA) for AAV production as described [36]. Briefly, recombinant self-complementary AAV vector plasmids containing the genes of interest were transfected into adherent HEK293 cells with AAV serotype 5 and Ad helper plasmids by CaPO4 transfection. Cell were fragmented and AAV particles were purified by iodixanol step gradient ultracentrifugation and ion exchange column chromatography [40,41]. AAV titers were determined by qPCR and purity was confirmed by SDS-PAGE as previously described [40,41].

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