title: A library of TAL effector nucleases spanning the human genome.
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.
authors: National Creative Initiatives Research Center for Genome Engineering and Department of Chemistry, Seoul National University, Gwanak-gu, Seoul, South Korea.
authors: Yongsub
authors: Y
authors: Kim
authors: Jiyeon
authors: J
authors: Kweon
authors: Annie
authors: A
authors: Kim
authors: Jae Kyung
authors: JK
authors: Chon
authors: Ji Yeon
authors: JY
authors: Yoo
authors: Hye Joo
authors: HJ
authors: Kim
authors: Sojung
authors: S
authors: Kim
authors: Choongil
authors: C
authors: Lee
authors: Euihwan
authors: E
authors: Jeong
authors: Eugene
authors: E
authors: Chung
authors: Doyoung
authors: D
authors: Kim
authors: Mi Seon
authors: MS
authors: Lee
authors: Eun Mi
authors: EM
authors: Go
authors: Hye Jung
authors: HJ
authors: Song
authors: Hwangbeom
authors: H
authors: Kim
authors: Namjin
authors: N
authors: Cho
authors: Duhee
authors: D
authors: Bang
authors: Seokjoong
authors: S
authors: Kim
authors: Jin-Soo
authors: JS
authors: Kim
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D030541:Databases, Genetic; D004720:Endonucleases / Q000235:genetics*; D017353:Gene Deletion; D055786:Gene Knockout Techniques; D015723:Gene Library; D005818:Genetic Engineering / Q000379:methods*; D015894:Genome, Human; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D016328:NF-kappa B; D015398:Signal Transduction; D014157:Transcription Factors
chemical_list: D016328:NF-kappa B; D014157:Transcription Factors; D004720:Endonucleases