Engineering a self-eliminating transgene in the yellow fever mosquito, Aedes aegypti

Autor: Keun Chae, Chanell Dawson, Collin Valentin, Bryan Contreras, Josef Zapletal, Kevin M Myles, Zach N Adelman
Rok vydání: 2022
Předmět:
Zdroj: PNAS Nexus. 1
ISSN: 2752-6542
DOI: 10.1093/pnasnexus/pgac037
Popis: 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.
Databáze: OpenAIRE