Engineering a biodegradable transgene in the dengue mosquito Aedes aegypti

On-Demand

Presenting Author(s)

Keun Chae

Texas A&M University
College Station, Texas

Co-Author(s)

CD

Chanell Dawson

Texas A&M University
College Station, Texas
CV

Collin Valentin

Texas A&M University
College Station, Texas
KM

Kevin Myles

Texas A&M University
College Station, Texas
ZA

Zach Adelman

Texas A&M University
College Station, Texas

Promising genetics-based approaches are being developed to reduce or prevent the transmission of mosquito-vectored diseases. Less clear is how such transgenes can be removed from the environment, a concern that is particularly relevant for highly invasive gene drive transgenes. Here, we lay the groundwork for a transgene removal system based on single-strand annealing (SSA), a eukaryotic DNA repair mechanism. A transgene we refer to as the SSA-based rescuer strain (kmo^RG) was engineered to have direct repeat sequences (DRs) in the Ae. aegypti kynurenine 3-monooxygenase (kmo) gene flanking the intervening cargo genes, DsRED and EGFP. Targeted induction of DNA double strand breaks (DSBs) in the DsRED transgene successfully triggered complete elimination of the entire cargo from the kmo^RG strain, restoring wild-type kmo gene and thereby normal eye pigmentation. Our work establishes the framework for strategies to remove transgene sequences during the evaluation and testing of modified strains for genetics-based mosquito control.