Table 1 Effect of CRISPR/Cas9 RNPs concentration on the screwworm survival, mutagenesis and germline transmission
Cas9 (ng/µl)sgRNA (ng/µl)Injected embryosHatched LarvaeaPupaeaAdult Survivors at G0abwb mosaics at G0bbwb transmission to G1 (1 ♂ G0mosaic x 2 ♀ bwb)d
TotalTotalFounderscInheritancee
UninjectedUninjected427323 (75.6%)157 (36.8%)76 (17.8%)67 (15.7%)143 (33.5%)000
MockMock320192 (60.0%)71 (22.2%)29 (9.1%)33 (10.3%)62 (19.4%)000
360145373180 (48.2%)82 (21.9%)29 (7.8%)34 (9.1%)63 (16.9%)15 (23.8%)20 (31.7%)35 (55.6%)8/10 (80%)38.4 ± 4.3%
500200400137 (34.2%)48 (12.0%)23 (5.75%)15 (3.75%)38 (9.5%)18 (47.4%)10 (26.3%)28 (73.7%)6/8 (75%)81.9 ± 14%
  • a, percentage relative to the number of embryos; b, percentage relative to the number of adult survivors; c, proportion of G0 bwb mosaic males that transmitted Cas9-derived mutant alleles to the G1 generation (founders) after crossing with bwb females from the ChYellow 07/01 mutant strain; d. crosses made to evaluate new mutated alleles for complementation allowing to estimate germline transmission; e, average ± SEM of G1 biallelic bwb mutants obtained from crossing G0 mosaic males and bwb females (individual result from each cross are shown in Table S3);