Figure 1

Sexing using female-specific splicing under the control of the repressible tTA-system. The depicted transgenic sexing system [41, 42] uses a sex-specifically spliced intron and a hyperactive pro-apoptotic gene to generate female-specific lethality under the regulation of the tetracycline-controlled transactivator (tTA). To cause early embryonic lethality and thus avoidance of larval survival, the tTA is under the control of an early embryonic promoter. During rearing of such strains, addition of tetracycline (TET) to the food keeps the system in the OFF state, as tetracycline blocks the binding of tTA to its response element (TRE). For the release generation, tetracycline is absent in the food and therefore the sexing system is ON: in males, the male specific splicing of the transformer intron (tra-I) - placed after the translation start codon (ATG) of the effector gene - includes a small exon containing a TAA stop codon between the start codon and the rest of the effector gene and therefore prevents the production of the functional pro-apoptotic effector protein allowing the males to survive; whereas in the females the female specific splicing of the tra-I produces a functional effector and the embryonic cells are driven into apoptosis, which leads to female-specific embryonic lethality.