Fig 2.

Comparison of AAV hybrid capsid genome packaging and in vitro transduction efficiencies. (A) Triple transfections of HEK293 cells were performed in 6-well plates, combining an AAV2 rep/AAV8-AAVrh32.33 hybrid cap packaging plasmid with a cis-plasmid containing CMV.ffLuc, AAV2 inverted terminal repeats, and an adenovirus helper construct (pAdΔF6). Titers of small-scale vector preparations are reported as the number of drp/ml (theoretically equivalent to the number of GCs/ml). All vectors were produced in a final volume of 3 ml. The data shown are means ± SDs from 3 wells within a 6-well plate. Results were confirmed in three separate experiments. (B and C) Small-scale vector preparations of AAV hybrid capsids encoding CMV.ffLuc were used to transduce HEK293 cells (B) or U937 human monocytes (C) in a 96-well plate at an MOI of 10⁴. Wild-type adenovirus helper was used for cotransduction at an MOI of 10². The data shown are means ± SDs from at least 3 duplicate wells on a 96-well plate. Results were confirmed in three separate experiments both with and without adenovirus helper. No Pack, a negative control in which no packaging plasmid was added to the transfection mixture; Cells Only, a negative control of untransfected cells.