Harvest of produced viral vectors
Viral particles were harvested from the cell culture broth as previously described [47, 56]. Briefly, the cells were harvested and lysed by adding a 10X Lysis Buffer (20 mM MgCl2, 1% Triton X-100 in 500 mM Tris-buffered solution). Benzonase was added to a final concentration of 5 U/mL to digest host-cell DNA and unpacked virus DNA. After 1 hour of incubation at 37°C with agitation, MgSO4was added to a final concentration of 37.5 mM to prevent AAV aggregation and binding to cellular components. After 30 minutes of incubation at 37°C with agitation, the lysate was clarified via centrifugation at 10,000 × g for 15 minutes. The clarified viral lysate was stored in a -80°C freezer.