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Abstract
Yongsub Kim1,5, Jiyeon Kweon1,5, Annie Kim1,5, Jae Kyung Chon2,5, Ji Yeon Yoo3,5, Hye Joo Kim1, Sojung Kim1, Choongil Lee1, Euihwan Jeong1, Eugene Chung1, Doyoung Kim1, Mi Seon Lee3, Eun Mi Go3, Hye Jung Song3, Hwangbeom Kim4, Namjin Cho4, Duhee Bang4, Seokjoong Kim3 & Jin-Soo Kim1
1National Creative Initiatives Research Center for Genome Engineering and Department of Chemistry, Seoul National University, Gwanak-gu, Seoul, South Korea. 2Department of Interdisciplinary Program in Bioinformatics, Seoul National University, Gwanak-gu, Seoul, South Korea. 3ToolGen, Inc., Geumcheon-Gu, Seoul, South Korea. 4Department of Chemistry, Yonsei University, Seoul, South Korea. 5These authors contributed equally to this work.
Correspondence to: Jin-Soo Kim or Seokjoong Kim
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.
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