Table 1.
TLR9 Activation Potential and Risk Factor (RF)* calculations for AAV genomes and components.
| DNA test article | Size (nt) | RF1 | RF3 | Normalized RF3 | TLR9 Activation |
|---|---|---|---|---|---|
| Human genome | |||||
| complete genome * | 3.21 x109 | 0.965 | 0.191 | 1.00 | - |
| Bacterial genome | |||||
| Escherichia coli * | 4.64 x106 | 7.471 | 4.683 | 24.5 | +++ |
| Clinical rAAV genomes | |||||
| low CpG AAV8-FIXsc (scAAV2/8-LP1-hFIXco42; clinicaltrials.gov NCT00979238) | 4611 | 1.757 | 1.298 | 6.80 | + |
| high CpG AAV8-FIXsc (BAX33534; clinicaltrials.gov NCT01687608) | 4780 | 5.859 | 4.383 | 22.9 | +++ |
| rAAV genome components | |||||
| AAV2 ITRs | 145 | 11.03 | 7.862 | 41.2 | +++ |
| ApoE HCR hAAT Promoter | 828 | 3.019 | 2.754 | 14.4 | +++ |
| MHCK7 Promoter | 770 | 2.078 | 1.851 | 9.69 | ++ |
| CAG Promoter | 584 | 8.562 | 2.603 | 13.6 | +++ |
| CMV Promoter | 508 | 5.709 | 3.740 | 19.6 | +++ |
| WPRE | 597 | 6.198 | 3.819 | 20.0 | +++ |
*Wright JF. Quantification of CpG motifs in rAAV genomes: avoiding the toll. Mol Ther 2020; 28(8):1756-58.
RF1 = f [CpGT / nt] X 100%.
RF3 = f [CpGT + CpGS4 – 2CpGI4 / nt] X f [CpGMeneg / CpGT ] X 100%.
Normalized RF3 = RF3 (test article) / RF3 (human genome).