Table 2.
Generation of founder knock-out lines by ends-out targeting
| Target gene | Screening cross progenya | Preliminary candidates | On-target chromosome | Genetically verified | PCR verified | HR frequencyb |
|---|---|---|---|---|---|---|
| DE-Cadc | ≈1.6 × 105 | ≈1,700 | 96/≈1,700 | 22/72d | 22/22 | ≈1.9 × 10−4 |
| lglc | ≈2.4 × 105 | 1,127 | 95/1,127 | 22/95e | 22 /22j | ≈9 × 10−5 |
| crumbsc | ≈1.8 × 105 | ≈400 | 26/≈400 | 1/26f | 1/1 | ≈6 × 10−6 |
| CG31158c | ≈1.5 × 105 | 1,140 | 8/1,140 | 1/8g | 1/1 | ≈7 × 10−6 |
| dArf6 | ≈7 × 105 | 315 | 30/315 | 5/30h | 5/5 | ≈7 × 10−6 |
| sdt | ≈1 × 106 | 116 | 4/116 | 4/4i | 4/4 | ≈4 × 10−6 |
aThe total estimated number of screening cross progeny (12) that were screened in each targeting experiment. Because all progeny were mixed and screened together, we did not register the clonality of the preliminary candidates. We assumed that each targeting mutant obtained was because of a distinct targeting event.
bBecause all female candidates (or male candidates as in the case of sdt targeting) were discarded in targeting experiments, the adjusted HR frequency should be twice as high as listed here.
cThe targeting constructs for these genes were made on an older version of pGX-attP that does not contain the UAS-Rpr (see SI Materials and Methods), hence the large numbers of false-positives among preliminary candidates.
dOnly 72 of 96 candidates were tested for noncomplementing the null allele shg2.
eAll candidates were tested for noncomplementing the null allele lgl4.
fAll candidates were tested for noncomplementing the null allele crb11A22.
gA previously generated knock-out allele, CG31158KO#1 (see Materials and Methods) was used for complementation assays.
hA dArf6ΔKG#1-deletion allele generated by P-excision (see Materials and Methods) was used for complementation assays.
iAll candidates were tested for noncomplementing the null allele sdtXP96 (24).
jSee Fig. S3 for details.