A library of TAL effector nucleases spanning the human genome

Yongsub Kim, Jiyeon Kweon, Annie Kim, Jae Kyung Chon, Ji Yeon Yoo, Hye Joo Kim, Sojung Kim, Choongil Lee, Euihwan Jeong, Eugene Chung, Doyoung Kim, Mi Seon Lee, Eun Mi Go, Hye Jung Song, Hwangbeom Kim, Namjin Cho, Duhee Bang, Seokjoong Kim, Jin Soo Kim

Research output: Contribution to journalArticlepeer-review

263 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)251-258
Number of pages8
JournalNature Biotechnology
Volume31
Issue number3
DOIs
Publication statusPublished - 2013 Mar

Bibliographical note

Funding Information:
This work was supported by the National Research Foundation of Korea (J.-S.K., 2012-0001225), the Intelligent Synthetic Biology Center of the Global Frontier Project funded by the Ministry of Education, Science and Technology, Korea (D.B., 2011-0031956), Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (S.K., 311062-04-2-sb1010), and Plant Molecular Breeding Center of Next-Generation BioGreen 21 Program (S.K., PJ009081).

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

Fingerprint

Dive into the research topics of 'A library of TAL effector nucleases spanning the human genome'. Together they form a unique fingerprint.

Cite this