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. 2008 Nov 21;19(12):1421–1427. doi: 10.1089/hum.2008.118

FIG. 2.

FIG. 2.

Use of a split Rep/Cap AAV helper system overcame the negative effects of the consensus donor, allowing for even greater levels of rAAV production. Left: Representative RNase protection assay of AAV2 CMV-driven capsid-encoding pre-mRNAs in the absence or presence of pHelper. Middle: Quantification of the relative spliced-to-unspliced ratios of capsid-encoding pre-mRNA. Data are from at least three experiments and show standard deviations. The position of the “RP” RNase protection probe is indicated. Right: Relative levels of rAAV production observed in AAV2, AAV6, and AAV8 with the “better” and consensus donor mutants in relation to levels obtained with the wild-type donors (set to a value of 1.00). Average titers of DNase-resistant rAAV virions per 100-mm dish (with standard deviations in parentheses) were as follows: CMV-Cap2, 1.95 × 1010 (±6.75 × 109); CMV-Cap2bD, 7.47 × 1010 (±1.78 × 1010); CMV-Cap2cD, 1.15 × 1011 (±4.82 × 1010); CMV-Cap6, 9.24 × 109 (±6.84 × 108); CMV-Cap6bD, 3.75 × 1010 (±2.41 × 109); CMV-Cap6cD, 5.45 × 1010 (±3.27 × 109); CMV-Cap8, 2.45 × 1010 (±7.27 × 109); CMV-Cap8bD, 8.59 × 1010 (±1.56 × 1010); CMV-Cap8cD, 1.25 × 1011 (±3.06 × 1010).