TABLE 1 .
IC50s for asexual-stage parasites of the indicated CRISPR/Cas9-engineered parasite lines for GNF179 and artemisinina
| Strain | Gene mutated (technique) | Mutation introduced | GNF179 IC50 (nM) |
Fold increase | Artemisinin IC50 (nM) (actual) | Fold increase | |
|---|---|---|---|---|---|---|---|
| Predicted (9) | Actual | ||||||
| Dd2 | NA | NA | 2.5 | 3.1 ± 0.9 | NA | 12 ± 0.8 | NA |
| Dd2 | pfcarl (CRISPR) | L830V | 16 | 61 ± 12 | 20 | 8.9 ± 0.5 | 0.74 |
| Dd2 | pfcarl (CRISPR) | S1076N | — | 140 ± 21 | 44 | 7.7 ± 0.7 | 0.64 |
| Dd2 | pfcarl (CRISPR) | S1076I | — | 110 ± 17 | 35 | 11 ± 1.7 | 0.92 |
| Dd2 | pfcarl (CRISPR) | V1103L | 62 | 83 ± 11 | 27 | 12 ± 0.2 | 1.0 |
| Dd2 | pfcarl (CRISPR) | I1139K | 2,669 | 1,400 ± 230 | 458 | 8.3 ± 1.3 | 0.69 |
| Dd2 | pfcarl (Bxb1) | OE | — | 6.4 ± 0.9 | 2.1 | 10 ± 2.3 | 0.84 |
| NF54 | NA | NA | 1.7 | 3.0 ± 1.7 | NA | 9.9 ± 1.4 | NA |
| NF54 | pfcarl (CRISPR) | L830V | — | 110 ± 23 | 35 | 8.3 ± 0.6 | 0.84 |
| NF54 | pfcarl (CRISPR) | S1076N | 48 | 83 ± 23 | 28 | 9.8 ± 0.9 | 0.99 |
| NF54 | pfcarl (CRISPR) | S1076I | — | 79 ± 7.0 | 26 | 8.1 ± 1.0 | 0.82 |
| NF54 | pfcarl (CRISPR) | V1103L | — | 53 ± 11 | 18 | 11 ± 1.1 | 1.1 |
| NF54 | pfcarl (Bxb1) | OE | — | 7.1 ± 1.0 | 2.4 | 8.5 ± 0.7 | 0.86 |
a
IC50s for the asexual-stage parasites were determined via the 72 h SYBR green I assay. Predicted IC50s are the values observed for drug-selected parasite lines harboring the indicated mutation, and a dash means that the corresponding specific mutation had never been observed in a single mutant in that strain and that the predicted values for NF54 were extrapolated from previously reported values for 3D7 (7). IC50s are listed as means ± SE (n = 3). NA, not applicable. OE, Over-expression.