Engineering a self-eliminating transgene in the yellow fever mosquito,Aedes aegypti
Abstract
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. An SSA-based rescuer strain (kmoRG) was engineered to have direct repeat sequences (DRs) in the Aedes aegypti kynurenine 3-monooxygenase (kmo) gene flanking the intervening transgenic 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 kmoRG strain, restoring the 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.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Mar 30, 2022
- Source ID
- 10.1093/pnasnexus/pgac037
Entities
People
- Bryan Contreras
- Chanell Dawson
- Collin Valentin
- Josef Zapletal
- Keun Chae
- Kevin M. Myles
- Zach Adelman
Organizations
- Defense Advanced Research Projects Agency
- National Institutes of Health
- Texas A&M University