FairyTALE: a high-throughput TAL effector synthesis platform.
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.
Department of Chemical and Biomolecular Engineering, ‡Institute for Genomic Biology, §Department of Biochemistry, University of Illinois at Urbana-Champaign , Urbana, Illinois 61801, United States of America.
Jing
J
Liang
Ran
R
Chao
Zhanar
Z
Abil
Zehua
Z
Bao
Huimin
H
Zhao
D055786:Gene Knockout Techniques; D005818:Genetic Engineering; D016680:Genome, Bacterial; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D058615:Synthetic Biology / Q000379:methods*; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014974:Xanthomonas / Q000235:genetics
D015534:Trans-Activators