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Transgene removal using an in cis programmed homing endonuclease via single-strand annealing in the mosquito Aedes aegypti

Commun Biol. 2024-05; 
Keun Chae, Bryan Contreras, Joseph S Romanowski, Chanell Dawson, Kevin M Myles, Zach N Adelman
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Gene Synthesis … DsRED-SV40 (AniI) contains the synthesized DNA piece (GenScript) that encodes DsRED with the homing endonuclease Y2-I-AniI recognition sequence was engineered in-frame next … Get A Quote

摘要

While gene drive strategies have been proposed to aid in the control of mosquito-borne diseases, additional genome engineering technologies may be required to establish a defined end-of-product-life timeline. We previously demonstrated that single-strand annealing (SSA) was sufficient to program the scarless elimination of a transgene while restoring a disrupted gene in the disease vector mosquito Aedes aegypti. Here, we extend these findings by establishing that complete transgene removal (four gene cassettes comprising ~8-kb) can be programmed in cis. Reducing the length of the direct repeat from 700-bp to 200-bp reduces, but does not eliminate, SSA activity. In contrast, increasing direct repeat length to 1.... More

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