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Nat Biotechnol. 2013 Mar;31(3):251-8. doi: 10.1038/nbt.2517. Epub 2013 Feb 17.

A library of TAL effector nucleases spanning the human genome.

Author information

1
National Creative Initiatives Research Center for Genome Engineering and Department of Chemistry, Seoul National University, Gwanak-gu, Seoul, South Korea.

Abstract

Transcription activator-like (TAL) effector nucleases (TALENs) can be readily engineered to bind specific genomic loci, enabling the introduction of precise genetic modifications such as gene knockouts and additions. Here we present a genome-scale collection of TALENs for efficient and scalable gene targeting in human cells. We chose target sites that did not have highly similar sequences elsewhere in the genome to avoid off-target mutations and assembled TALEN plasmids for 18,740 protein-coding genes using a high-throughput Golden-Gate cloning system. A pilot test involving 124 genes showed that all TALENs were active and disrupted their target genes at high frequencies, although two of these TALENs became active only after their target sites were partially demethylated using an inhibitor of DNA methyltransferase. We used our TALEN library to generate single- and double-gene-knockout cells in which NF-κB signaling pathways were disrupted. Compared with cells treated with short interfering RNAs, these cells showed unambiguous suppression of signal transduction.

PMID:
23417094
DOI:
10.1038/nbt.2517
[Indexed for MEDLINE]

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