Abstract
Recombinant transcription activator-like effectors (TALEs) have been effectively used for genome editing and gene regulation applications. Due to their remarkable modularity, TALEs can be tailored to specifically target almost any user-defined DNA sequences. Here, we introduce fairyTALE, a liquid phase high-throughput TALE synthesis platform capable of producing TALE-nucleases, activators, and repressors that recognize DNA sequences between 14 and 31 bp. It features a highly efficient reaction scheme, a flexible functionalization platform, and fully automated robotic liquid handling that enable the production of hundreds of expression-ready TALEs within a single day with over 98% assembly efficiency at a material cost of just $5 per TALE. As proof of concept, we synthesized and tested 90 TALEs, each recognizing 27 bp, without restrictions on their sequence composition. 96% of these TALEs were found to be functional, while sequencing confirmation revealed that the nonfunctional constructs were all correctly assembled.
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