Location: Insect Behavior and Biocontrol Research
Project Number: 6036-22000-030-01-N
Project Type: Non-Funded Cooperative Agreement
Start Date: Sep 30, 2014
End Date: Sep 29, 2019
The objective of this cooperative research project is to develop mechanisms for conditional female-specific lethality in Bactrocera species to create transgenic strains that will improve the sterile insect technique (SIT) for the control of these pest populations. Specific goals of this agreement will be to determine the molecular mechanisms of female-specific splicing of sex determination gene introns and to test lethal effector genes. These components will be used together in a tetracycline-suppressible system that allows, in the absence of the antibiotic, lethal effector gene expression solely in females for female-specific lethality.
PCR approaches will be taken to identify and isolate the transformer and doublesex sex determination genes from the Bactrocera species of interest using homologous sequences from B. dorsalis for primer design. Female-specific introns from the sex determination genes will then be incoporated into the Tet-transactivator (TTA) driver cassette. The hid and reaper pro-apoptotic cell death genes will be similarly isolated from the Bactrocera species, and their coding sequences will be linked in-frame to the Tet-response element (TRE) to create lethal effector cassettes. Both the driver and effector cassettes will be incorporated into piggyBac vectors and transformed into the host Bactrocera species. Progeny from appropriate matings between driver and lethal effector transformant lines will then be tested on Tet-containing and Tet-free diet to evaluate female-specific lethality in order to produce male-only lines for sterile male releases.