AAV vectors

Recombinant adeno-associated viral vectors of serotype 6 (AAV-6) were prepared by transient transfection of vector genome plasmids (for details see Supplemental Figure 1) with the DP6 helper plasmid in HEK293 cells, viral particles were purified from cell lysates by iodixanol gradient centrifugation and heparin affinity chromatography. After extensive dialysis against PBS particles were frozen in single use aliquots at −80°C. Genome titres were determined by qPCR and >98% purity was confirmed by SDS-PAGE. Vector genome (vg) titre vs. transducing units (tu) titre was determined with EGFP expressing vectors in primary neurons and was estimated to be 1:30 (tu:vg). All constructs expressed transgenes from a neuron-specific synapsin1 gene promoter.