期刊
NATURE BIOTECHNOLOGY
卷 30, 期 5, 页码 460-+出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nbt.2170
关键词
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资金
- US National Institutes of Health (NIH) [DP1 OD006862]
- NIH [P50HG005550, T32 CA009216]
- Jim and Ann Orr MGH Research Scholar Award
- National Science Foundation [DBI-0923827]
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.
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