Fertility Evaluation of Germline Gene Silencing

To test how silencing of a candidate gene in the germline affects fertility, a test vial was set up 10 days after crossing the respective RNAi fly stock with the nos-GAL4-VP16 MVD1 driver. In this vial, five females and three males from the non-balanced F1 progeny were incubated for 15 days at 25 °C before recording fertility. The F1 progeny was considered to be fertile when the number of progeny in the F2 generation was comparable to wild-type flies. When the number of F2 progeny was substantially reduced, the F1 progeny was considered to have reduced fertility. The F1 progeny was considered sterile in the complete absence of larvae or pupae.