title: TALEN-based knockout library for human microRNAs.
abstract: Various technical tools have been developed to probe the functions of microRNAs (miRNAs), yet their application has been limited by low efficacy and specificity. To overcome the limitations, we used transcription activator-like effector nucleases (TALENs) to knock out human miRNA genes. We designed and produced a library of 540 pairs of TALENs for 274 miRNA loci, focusing on potentially important miRNAs. The knockout procedure takes only 2-4 weeks and can be applied to any cell type. As a case study, we generated knockout cells for two related miRNAs, miR-141 and miR-200c, which belong to the highly conserved miR-200 family. Interestingly, miR-141 and miR-200c, despite their overall similarity, suppress largely nonoverlapping groups of targets, thus suggesting that functional miRNA-target interaction requires strict seed-pairing. Our study illustrates the potency of TALEN technology and provides useful resources for miRNA research.
authors.affiliation: 1] Center for RNA Research, Institute for Basic Science, Seoul, Korea. [2] School of Biological Sciences, Seoul National University, Seoul, Korea. [3].
authors.forename: Young-Kook
authors.initials: YK
authors.lastname: Kim
authors.forename: Gabbine
authors.initials: G
authors.lastname: Wee
authors.forename: Joha
authors.initials: J
authors.lastname: Park
authors.forename: Jongkyu
authors.initials: J
authors.lastname: Kim
authors.forename: Daehyun
authors.initials: D
authors.lastname: Baek
authors.forename: Jin-Soo
authors.initials: JS
authors.lastname: Kim
authors.forename: V Narry
authors.initials: VN
authors.lastname: Kim
mesh_terms: D001483:Base Sequence; D055786:Gene Knockout Techniques; D015723:Gene Library; D006801:Humans; D035683:MicroRNAs / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012260:Ribonucleases / Q000737:chemistry* / Q000235:genetics; D017421:Sequence Analysis
chemical_list: C533896:MIRN200 microRNA, human; D035683:MicroRNAs; D012260:Ribonucleases