Fig. 4: Functionality of SGyA as a split gene drive using a re-coded GDe.

a The crossing schematic involves the paternal Cas9 from the Y chromosome. The Cas9 transgene is only passed through the male germline. A SGyA male is crossed to homozygous GDe females to produce transheterozygous males and GDe-only females. The F1 transheterozygous male is then outcrossed to a w- females to assess GDe inheritance in F2 progeny. b The crossing schematic involves the paternal Cas9 from an autosome. A homozygous male harboring Cas9 on an autosome is outcrossed to a homozygous GDe female. Transheterozygous F1 males are subsequently outcrossed to w- females to assess GDe inheritance. Both sexes are affected. c Crossing schematic of the negative control cross. d Percent of F2 eggs hatched in crosses involving the control, transheterozygous SGyA males and transheterozygous autosomal Cas9 males. e Inheritance of the GDe in F2 progeny among different sources of Cas9. f Inheritance of the GDe in F2 and F3 progeny from crosses involving a transheterozygote male containing the GDe and the SGyA transgene. No significant deviations between both the F2 and F3 data sets were found (p value = 0.3446). g Inheritance of the GDe in F2 and F3 progeny from crosses involving a transheterozygote male containing the GDe and the autosomal Cas9 transgene. Significant differences between GDe inheritance observed in F2 progeny (when compared to control; p value = 0.0129) and GDe inheritance between F2 and F3 progeny (p value = 0.0051) Blue shaded boxes in crossing schematics highlight instances where a bias of GDe transmission is observed. Green arrows indicate the conversion of WT PolG2 allele into the GDe. Gray numbers represent the expected Mendelian inheritance percentages of the GDe. Green percentages indicate the homing/bias of GDe. At least 18 experimental/replicate crosses were set up per Cas9 experiment, and 6 for the control to determine F2 progeny outcomes. For F3 outcomes, at least nine experimental crosses were performed. Significance was determined using a two-tailed unpaired student’s t test. For inheritance and male bias plots, vertical bars represent SEM. n.s. represents Non-significant. ***p < 0.0005; **p < 0.001; *p < 0.01. Source data is provided as a Source Data file.