Young-Kook Kim1,2,6, Gabbine Wee3,4,6, Joha Park1,2, Jongkyu Kim1,2, Daehyun Baek1,2,5, Jin-Soo Kim3,4 & V Narry Kim1,2
1Center for RNA Research, Institute for Basic Science, Seoul, Korea. 2School of Biological Sciences, Seoul National University, Seoul, Korea. 3National Creative Research Initiatives Center for Genome Engineering, Seoul National University, Seoul, Korea. 4Department of Chemistry, Seoul National University, Seoul, Korea. 5Bioinformatics Institute, Seoul National University, Seoul, Korea. 6These authors contributed equally to this work.
Correspondence to: V Narry Kim or Jin-Soo Kim
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.