FairyTALE: a high-throughput TAL effector synthesis platform

ACS Synth Biol. 2014 Feb 21;3(2):67-73. doi: 10.1021/sb400109p. Epub 2013 Nov 22.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Gene Knockout Techniques
  • Genetic Engineering
  • Genome, Bacterial
  • Plasmids / genetics
  • Plasmids / metabolism
  • Synthetic Biology / methods*
  • Trans-Activators / genetics
  • Trans-Activators / metabolism*
  • Xanthomonas / genetics

Substances

  • Trans-Activators