Fig. 7: Cas12a-based pgSIT males reduce female reproductive output at 25 °C.

a Initial sterility and lethality assessment. Transhomozygous males from the triple-transgene line were crossed with WT females at 25 °C. While female lethality was previously complete at 29 °C, partial escapees (~9%) were observed at 25 °C. Single-pair crosses with 38 pgSIT males revealed that only 2 were fertile, indicating strong sterility. Three independent experiments from the G0 (n = 3) were performed to then collect single males to test sterility at the G1 level. Data are presented as mean values +/− SD. More details are provided in Source Data File 7. b Cage assays were performed with 10 WT females and 4 males under three conditions. Adults mated for 48 h before removal. After parents removal, all pgSIT males recovered from “mixed” and pgSIT-only cages were confirmed sterile by single-pair crosses with virgin WT females (Source Data File 8). Created in BioRender. Lopez del Amo, V. (2025) https://BioRender.com/gevpi2n. c At least, 3 biological replicates (n = 3) were evaluated for each condition. Data are presented as mean values +/− SD. Control cages averaged 175 eggs, while mixed cages averaged 120 eggs (32% reduction). The pgSIT-only condition yielded almost no eggs. Hatching rates dropped from 48% (control) to 37% (mixed), with none in pgSIT-only. ANOVA and Tukey’s multiple comparisons test were performed for statistical analysis (see details in Source Data File 8).