Project Number: 2040-22430-027-034-I
Project Type: Interagency Reimbursable Agreement
Start Date: Oct 1, 2017
End Date: Sep 30, 2022
1. Screen the genomes of existing medfly genetic sexing strains to characterize the mutations causing the temperature sensitive lethal and white pupae traits. 2. Characterize the exact translocation breakpoint between the Y chromosome and the 5th autosome in the males of the Veinna-8 strain that confers these traits to be female specific. 3. Adapt CRISPR-based methods to recreate the translocation in wild type lines. 4. Demonstrate a targeted gene editing approach to re-creating temperature. sensitive lethal in Anastrepha and Bactrocera species.
The overall concept of this proposal is to first characterize the mechanism of the existing genetic sexing system in the medfly Vienna-8 strain using forward genetic techniques, validate those methods through reverse genetic screens using targeted gene editing techniques, and then re-create orthologous mutations in wild type lines of Bactrocera and Anastrepha for which robust genetic sexing strains do not currently exist. This is outlined in the following objectives.