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- Title
FLASH assembly of TALENs for high-throughput genome editing.
- Authors
Reyon, Deepak; Tsai, Shengdar Q; Khayter, Cyd; Foden, Jennifer A; Sander, Jeffry D; Joung, J Keith
- Abstract
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.
- Subjects
GENETIC transcription; GENETIC engineering; GENOMES; CANCER genetics; EPIGENESIS
- Publication
Nature Biotechnology, 2012, Vol 30, Issue 5, p460
- ISSN
1087-0156
- Publication type
Article
- DOI
10.1038/nbt.2170