Table 3 CRISPR screening results
IDInjected FliesaDonorLarvaeFounder Fertilityb
MaleFemale
1vas-Cas9.RFP(−) (/FM7a,Tb)rnCR-EGFP17320(2)/44(3)22(6)/59(10)
2vas-Cas9.RFP(−) (/FM7a,Tb)rnCR-FLAG15117(2)/49(3)22(4)/37(6)
3vas-Cas9.RFP(−); roe[3]/TM3rnCR-EGFP1888/3118/42
4vas-Cas9.RFP(−); roe[3]/TM3rnCR-FLAG1583/4316/40
5vas-Cas9.RFP(−); rn-EGFP DsRed(−)rnC-STOP17421/6015/64
Founders Yielding DsRed+ G1cFounders with DsRed+ Offspring Yielding Targeted Tagging EventdOverall Germline Transmissione
MaleFemaleMaleFemaleMaleFemale
9 (2)/20 (2)2/22 (6)3 (1)/9 (2)1/23 (1)/20 (2); 15.0%1/22 (6); 4.5%
6 (1)/17 (2)4/22 (4)4 (1)/6 (1)3/44 (1)/17 (2); 23.5%3/22 (4); 13.6%
1/81/180/10/10/8; 0%0/18; 0%
0/33/160/01/30/3; 0%1/16; 6.3%
13/217/1511/137/711/21; 52.4%7/15; 46.7%
  • a vas-Cas9.RFP(−) chromosome is from Bloomington stock #55821; roe[3] chromosome is from #7411. The majority of the injected embryos from ID 1 and 2 are homozygous for vas-Cas9.RFP(−); all the rest are homozygous for vas-Cas9.RFP(−).

  • b The format is the number of fertile flies over the number of survived adult flies. The numbers in parentheses are for founders of vas-Cas9.RFP(−)/FM7a,Tb genotype.

  • c The format is the number of flies in the category over the number of fertile flies. The numbers in parentheses are for founders of vas-Cas9.RFP(−)/FM7a,Tb genotype. At least 60 or all G1 flies from each fertile founder were screened for DsRed.

  • d The format is the number of flies in the category over the number of founders yielding DsRed+ G1. The numbers in parentheses are for founders of vas-Cas9.RFP(−)/FM7a,Tb genotype. At least 60 or all G1 flies from each fertile founder were screened for DsRed, and approximately 10 individual DsRed+ G1 (unless fewer flies were recovered, which would all be used) from each candidate founder were crossed. For ID 1 and 2, all fertile DsRed+ G1 were PCR-screened and stained to test the presence of tags. For each founder lineage, at least one G1 fly with positive results for all the tests was sequenced for the targeted region to confirm a clean homologous recombination event. For ID 3, 4, and 5, G2 larvae were stained to check the tags before sequencing to confirm targeted events. Potential off-targets regions were PCR-amplified for sequencing.

  • e The percentage is calculated as the proportion of fertile founders that yield targeted tagging events. The numbers in parentheses are for founders of vas-Cas9.RFP(−)/FM7a,Tb genotype.