FLASH Assembly of TALENs Enables High-Throughput Genome Editing

By Deepak Reyon, S. Tsai, Cyd Khayter, Jennifer A. Foden, Jeffry D. Sander, J. Keith Joung. Nature Biotechnology, 2012

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Engineered transcription activator–like effector nucleases (TALENs) have shown promise as facile and broadly applicable genome editing tools. However, no publicly available high-throughput method for constructing TALENs has been published, and large-scale assessments of the success rate and targeting range of the technology remain lacking. Here we describe the fast ligation-based automatable solid-phase high-throughput (FLASH) system, a rapid and cost-effective method for large-scale assembly of TALENs. We tested 48 FLASH-assembled TALEN pairs in a human cell–based EGFP reporter system and found that all 48 possessed efficient gene-modification activities. We also used FLASH to assemble TALENs for 96 endogenous human genes implicated in cancer and/or epigenetic regulation and found that 84 pairs were able to efficiently introduce targeted alterations. Our results establish the robustness of TALEN technology and demonstrate that FLASH facilitates high-throughput genome editing at a scale not currently possible with other genome modification technologies.

Subfields: Molecular Biology, Plant Science

View Paper: https://www.semanticscholar.org/paper/81aeedea79b3ba4272b4188d9ebc907e658f43e9

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Citations in the corpus

TALENs: a widely applicable technology for targeted genome editing (2012, positive): “Although the high success rate and robust targeting range of dimeric TALENs has been recently established, 18 relatively less is known about these parameters for monomeric engineered TALE-based transcriptional activators or repressors.”
Transcriptional enhancers: from properties to genome-wide predictions (2014, methodology): “In addition to the functional manipulation of enhancers, related methods based on TALE nucleases (TALENs) 140 and CRISPR–Cas9 141,142 also allow the direct editing of the genomic DNA sequence.”