Effect of the rep gene sequences on AAV genome replication in MRN-negative or -positive cells. (A) Structure of the wt and recombinant AAV genomes used. The AAVtCR genome is composed of the p5 promoter and the rep ORF fused at its N terminus to the TAP-Cherry sequences (14). The AAVp5GFP vector contains the entire p5 promoter (37). (B) HeLa shRad50 and HeLa shLuc cells were coinfected with AAVtCR (MOI, 103 GP/cell) and either HSVΔPol or wt HSV (MOI, 5 PFU/cell). DNA extracted 20 h p.i. was analyzed by qPCR using rep-specific primers. *, P < 0.05. (C) HeLa shRad50 and HeLa shLuc cells were coinfected with either AAVGFP or AAVp5GFP (MOI, 103 GP/cell) and HSVΔPol (MOI, 5 PFU/cell). Rep activity was provided either by transfection of pRep or coinfection with wt AAV. GFP DNA levels were measured by qPCR.