Table 1 Germline transmission rates for mutations in e and lbk
(A) No. of Fertile PNo. (%) of Broods from P Crosses Without eNo. (%) of Broods from P Crosses with eNo. of F1 Lines Sequenced
6826 (38)42 (62)120
nos-Cas9 embryos were injected with a mix of gRNA-e, gRNA-lbk1, and gRNA-lbk2. P indicates the number of injected, fertile flies used in Parental crosses. The rate of e mutations: of the 68 P, 38% produced no ebony offspring, and 62% produced ebony offspring, with individual broods ranging from 1 to 100% ebony. 120 F1-derived balanced lines were sequenced. 235 sequences of the two lbk genomic regions were analyzed.
(B) % ebony in F1 BroodsNo. of Lines Sequenced for lbk-1No. (%) of Lines with lbk-1 MutationsNo. of Lines Sequenced for lbk-2No. (%) of Lines with lbk-2 MutationsNo. (%) of Lines with Both lbk-1 and lbk-2 Mutations
0240 (0)240 (0)0 (0)
1–505412 (22)5125 (49)10 (20)
>504222 (52)4028 (70)14 (36)
  • nos-Cas9 embryos were injected with a mix of gRNA-e, gRNA-lbk1, and gRNA-lbk2. Each F1 brood is grouped by percentage of ebony: 0, 1–50% low, and >50% jackpot. Performing a two-tailed Fisher’s Exact test (2 × 2 contingency table), we show that for lbk-1, there is a strong statistical significance when comparing the 0% ebony group to either the 1–50% ebony (P < 0.05) or the >50% ebony group (P < 0.0001) for presence of mutations at lbk-1. Additionally, there is a significant increase in the number of mutations at lbk-1 when comparing the 1–50% to >50% ebony group (P < 0.005). Similarly, for lbk-2, we show that there is a strong statistical significance when comparing the 0% ebony group to either the 1–50% (P < 0.0001) or the >50% ebony group (P < 0.0001) for presence of mutations at lbk-2.