10.1371/journal.pone.0063074.g004 Ruozhen Hu Ruozhen Hu Jared Wallace Jared Wallace Timothy J. Dahlem Timothy J. Dahlem David Jonah Grunwald David Jonah Grunwald Ryan M. O'Connell Ryan M. O'Connell Using two TALEN pairs to delete the entire human miR-155 hairpin sequence. Public Library of Science 2013 Biochemistry Nucleic acids rna RNA interference biotechnology Genetic engineering Computational biology genomics Functional genomics Molecular genetics Gene regulation gene expression genetics Genetic mutation mutagenesis Gene function Molecular cell biology talen pairs delete mir-155 hairpin 2013-05-07 02:31:46 Figure https://plos.figshare.com/articles/figure/_Using_two_TALEN_pairs_to_delete_the_entire_human_miR_155_hairpin_sequence_/699106 <p>(A) TALEN pairs targeting miR-155 were designed and constructed (called TALEN C). The upper panel shows a schematic of the TALEN A pair binding sites. The lower panel shows the sequence alignments comparing Wt and TALEN C mutated miR-155. The left and right TALEN binding sites are highlighted in yellow and the miR-155 seed region is boxed in red. (B) Schematic of the binding sites of two TALEN pairs (TALEN A and TALEN C) targeting miR-155. (C–D) Both TALEN A and TALEN C pairs were transfected into 293T cells. The miR-155 locus was amplified by PCR and subjected to TOPO cloning and sequencing. (C) Electrophoresis gel analysis showing deletions in the miR-155 locus. The arrows on the right indicate the two expected PCR products with or without large deletions. (D) Sequence alignments between a Wt clone and two TOPO clones with large deletions. The left and right TALEN binding sites for both TALEN A and TALEN C are highlighted in yellow and the miR-155 hairpin sequence is boxed in red.</p>