Figure 3. Population suppression in multigenerational population cage experiments.

(A) Multiple precision-guided sterile insect technique (pgSIT):wild-type (WT) release ratios, such as 1:1, 5:1, 10:1, 20:1, and 40:1, were tested in triplicate. A schematic diagram depicts the cage experiments for the 5:1 ratio. To start the first generation (G1) of multigenerational population cages, 250 mature pgSIT adult ♂’s were released with 50 similarly aged WT adult ♂’s into a cage, and in 1 hr 50 virgin ♀’s were added. The mosquitoes were allowed to mate for 2 days before all ♂’s were removed, and ♀’s were blood fed and eggs were collected. All eggs were counted, and 100 eggs were selected randomly to seed the next generation (G2). The sex-sorted adult ♂’s that emerged from 100 eggs (N_♂) were released with five times more (5N_♂) pgSIT adult ♂’s into a cage, and N_♀ ♀’s were added later. The remaining eggs were hatched to measure hatching rates and score transgene markers. This procedure was repeated for five generations for each cage population (Supplementary file 1i). Two types of pgSIT ♂’s were assessed: those that inherited a maternal Cas9 (pgSIT^♀Cas9), and those that inherited a paternal Cas9 (pgSIT^♂Cas9). Multigeneration population cage data for each release ratio plotting the normalized percentage of laid eggs (B) and hatched eggs (C). Points show means ± standard deviation (SD) for each triplicate sample (n=3). Source data are provided in Supplementary file 1.