TABLE 2.
Optimization experiment performed to optimize the editing of mRNA by GST-APOBEC1 and establish response surfaces
| Sample | pH | Temperature °C | Ionic strength mM | Divalent ion mM | −ACF editinga% | +ACF editinga % |
|---|---|---|---|---|---|---|
| 37 | 6.9 | 50 | 92.92 | 18 | 19.0 ± 2.8 | 14.3 (1) |
| 38 | 7 | 50 | 65 | 6 | 20.7 ± 1.0 | 14.7 (1) |
| 39 | 7.5 | 50 | 105 | 18 | 19.8 ± 4.2 | 8.93 ± 0 |
| 40 | 7.5 | 50 | 81.99 | 18 | 23.9 ± 1.0 | ND |
| 41 | 7.1 | 50 | 105 | 6 | 22.6 ± 3.5 | ND |
| 42 | 6.5 | 50 | 105 | 18 | 20.8 ± 6.8 | ND |
| 43 | 6.5 | 50 | 85.17 | 11.03 | 26.2 ± 7.2 | ND |
| 44 | 6.9 | 60 | 92.92 | 18 | 8.0 ± 0.8 | ND |
| 45 | 7 | 60 | 65 | 6 | 14.9 ± 4.2 | ND |
| 46 | 7.5 | 60 | 105 | 18 | 8.1 ± 3.6 | ND |
| 47 | 7.5 | 60 | 81.99 | 12.05 | 10.5 ± 1.9 | ND |
| 48 | 7.1 | 60 | 105 | 6 | 11.7 ± 3.3 | ND |
| 49 | 6.5 | 60 | 105 | 18 | 11.0 ± 1.0 | 9.6 (1) |
| 50 | 6.5 | 60 | 85.17 | 11.3 | 14.0 ± 1.0 | ND |
| 51 | 5.9 | 60 | 25 | 18 | 3.5 ± 4.4 | 1.8 ± 0.8 |
| 52 | 6.9 | 42 | 92.92 | 9.89 | 20.6 ± 1.8 | 23.2 ± 34 |
| 53 | 6.5 | 35 | 65 | 6 | 11.7 ± 1.7 | 27.0 ± 3.6 |
| 54 | 7.5 | 35 | 77.96 | 12.02 | 9.3 ± 1.7 | 18.3 ± 2.2 |
| 55 | 7.1 | 35 | 105 | 18 | 11.3 ± 2.5 | 17.2 ± 0.9 |
| 56 | 7.5 | 28 | 99.84 | 10.59 | 5.4 ± 1.3 | 13.8 ± 0.3 |
| 57 | 7.1 | 28 | 73.77 | 12.54 | 6.8 ± 0.28 | 15.3 ± 1.1 |
| 58 | 7 | 35 | 86.81 | 6 | 11.8 ± 1.7 | 14.9 ± 0.3 |
| 59 | 7.5 | 28 | 105 | 18 | 4.6 ± 0.3 | 14.8 ± 1.3 |
| 60 | 6.5 | 28 | 87.35 | 6 | 6.7 ± 0.2 | 15.0 ± 0.5 |
| 61 | 7 | 42 | 65 | 6 | 18.7 ± 0.5 | 25.6 ± 4.9 |
| 62 | 6.5 | 33 | 102.52 | 18 | 10.7 ± 1.3 | 22.3 ± 3.6 |
| 63 | 7.5 | 42 | 105 | 18 | 19.4 ± 0.6 | 11.0 ± 2.8 |
| 64 | 6.9 | 35 | 93.1 | 6 | 12.7 ± 1.1 | 16.4 ± 1.1 |
| 65 | 6.9 | 28 | 81.28 | 12.05 | 7.2 ± 1.3 | 20.6 (1) |
| 66 | 7.5 | 42 | 81.99 | 6 | 18.5 ± 2.0 | 17.8 ± 0.9 |
| 67 | 7.1 | 42 | 105 | 18 | 16.0 ± 2.1 | 16.5 ± 1.6 |
| 68 | 6.5 | 42 | 105 | 17.26 | 19.4 ± 0.9 | 21.9 ± 1.4 |
| 69 | 7.5 | 33 | 97.61 | 11.03 | 12.2 ± 0.3 | 15.1 ± 0.5 |
| 70 | 6.5 | 42 | 85.17 | 15.54 | 18.8 ± 0.2 | 24.5 ± 0.1 |
| 71 | 6.5 | 28 | 105 | 18 | 6.9 ± 0.7 | 18.1 ± 1.3 |
| 72 | 5.5 | 49 | 25 | 9.89 | 4.7 ± 1.6 | 3.3 ± 0.3 |
| 73 | 6.5 | 49 | 57.4 | 6 | 19.0 ± 1.4 | 11.4 ± 3.0 |
| 74 | 6.1 | 47 | 125 | 12.02 | 14.2 ± 3.2 | 10.2 ± 1.6 |
| 75 | 6.5 | 36 | 104.53 | 18 | 10.7 ± 3.6 | 17.3 ± 0.9 |
| 76 | 6.1 | 36 | 25 | 10.59 | 9.8 ± 1.8 | 17.3 ± 1.3 |
| 77 | 6.0 | 48 | 79.53 | 12.54 | 12.7 ± 1.7 | 8.1 ± 0.3 |
| 78 | 6.5 | 36 | 125 | 6 | 7.4 ± 0.4 | 17.9 ± 1.8 |
| 79 | 5.5 | 36 | 25 | 18 | 4.1 ± 1.1 | 8.3 ± 1.4 |
| 80 | 6.0 | 60 | 25 | 6 | 3.3 ± 0.5 | 4.4 ± 0.3 |
| 81 | 5.5 | 44 | 118.81 | 6 | 5.9 ± 1.2 | 4.0 ± 1.3 |
| 82 | 5.0 | 60 | 125 | 18 | 2.6 ± 0.3 | 1.9 ± 0.8 |
| 83 | 5.9 | 47 | 75.17 | 18 | 13.6 ± 0.14 | 7.6 ± 1.3 |
| 84 | 5.9 | 36 | 63.21 | 6 | 10.5 ± 2.8 | 13.8 ± 2.0 |
| 85 | 6.5 | 60 | 66.94 | 12.05 | 7.4 ± 1.3 | 6.9 ± 0.9 |
| 86 | 6.1 | 60 | 125 | 6 | 5.3 ± 1.1 | 4.7 ± 2.0 |
| 87 | 5.5 | 60 | 125 | 18 | 2.9 ± 0.02 | 2.7 ± 0.6 |
| 88 | 6.5 | 44 | 25 | 17.26 | 17.4 ± 2.5 | 17.8 ± 0.5 |
| 89 | 5.5 | 60 | 75.42 | 11.03 | 5.2 ± 3.1 | 2.6 ± 1.5 |
| 90 | 5.5 | 36 | 125 | 15.54 | 6.3 ± 0.3 | 8.84 ± 0.3 |
| 91 | 5.0 | 49 | 104.53 | 18 | 3.0 ± 0.7 | 2.2 ± 1.9 |
| 92 | 5.0 | 36 | 125 | 9.89 | 2.9 ± 1.4 | 3.2 ± 1.6 |
| 93 | 5.0 | 60 | 66.94 | 6 | 3.1 ± 1.2 | 2.9 ± 1.3 |
| 94 | 5.0 | 44 | 25 | 12.02 | 2.7 ± 0.9 | 2.0 ± 1.1 |
aPercentage editing activity is expressed as an average; n = 2–5, except where noted (1).